AB199091

Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control

RabMAb
Recombinant
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(33 Publications)

Anti-Rabbit IgG, monoclonal [EPR25A] - Isotype Control- Alexa Fluor® 488 conjugated (ab199091) is a rabbit recombinant monoclonal antibody used in Flow Cytometry, ICC/IF.

- Biophysical QC for unrivalled batch-batch consistency
- Over 20 publications

41 Images

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

ICC/IF

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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199091 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (pseudocolored in red). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD300a with ab325226 at 1/500 dilution (0.1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Cells were co-stained with anti human CD14 conjugated to Brilliant Violet 421.
Gated on viable cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD11a with ab317289 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Gated on viable cells. Cells are co-stained with CD3 conjugated to Alexa Fluor® 647.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling CD300a with ab325226 at 1/50 dilution (1ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Gated on viable cells.
Negative control : 293T (PMID : 22043923)

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing wild-type HAP1 (green line) and VDAC1 knockout HAP1 stained with ab317974 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab317974) (1x 10⁶ in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.

Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HAP1 WT cells (black line) and HAP1-VDAC1 KO cells (grey line), used at the same concentration and conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Cells were co-stained with anti human CD19 conjugated to PE/CY7.
Gated on viable cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) cells labelling TREM1 with ab316941 at 1/50 dilution (1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Human PBMC are co-stained with CD11b conjugated BV421.
Gated on viable cell.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab326161 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab326161) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing left wild-type HEK293T positive cells and right negative MFN2 knockout HEK293T stained with ab326161 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326161) (1x 106 in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C. Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab196467 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab196467) (1x 10⁶ in 100μl at 1 μg/ml (1/500)) for 30min on ice. Isotype control antibody was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 μg/ml PHA for 18h (bottom), either stained with ab309660 (right) or isotype control (left). The cells were fixed using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab309660) (1x 10⁶ in 100μl at 1 μg/ml (1/200)) for 30min on ice. Isotype control antibody was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab184818 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab184818) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing human peripheral mononuclear cells (PBMCs), either stained with ab317974 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab317974) (1x 10⁶ in 100μl at 0.2 μg/ml (1/2500)) for 30min on ice. Isotype control antibody was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control. The cells were simultaneously stained with CD3.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with myc-tagged IL-23R expression vector containing a Myc tag (Middle) / 293T cells transfected with an empty vector containing a Myc tag (Right) cells labelling IL-23R with ab319083 at 1/5000 dilution (0.01ug)compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody) (Blue).

Live cells were surface stained with ab319083, then fixed with 2% PFA followed by intracellular staining with Myc-tag conjugated to Alexa Fluor® 647.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling TCR delta with ab317064 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Gated on viable cells. Cells were co-stained with anti-CD4 conjugated to Alexa Fluor® 647.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing wild-type Jurkat (green line) and IDH2 knockout Jurkat stained with ab326446 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326446) (1x 106 in 100μl at 0.04 μg/ml (1/12500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in Jurkat WT cells (black line) and Jurkat-IDH2 KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.This antibody gave a positive signal in wild-type Jurkat fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt

Lab

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling Vimentin (phospho S56) with ab312898 at 1/500 dilution (0.1 µg)/Red (Red) compared with a Rabbit monoclonal IgG (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Cells were co-stained with DRAQ5 to differentiate cell cycle phase.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing HeLa cells stained with ab303582 (red line). The cells were fixed with 80 % methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were incubated in 1x PBS containing 10 % normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab303582) (1x 106 in 100μl at 0.2μg/ml (1/2500)) for 30 min at 22°C.

Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor 488 ® (ab199091) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488 nm) and 525/40 bandpass filter.

This antibody gave a positive signal in HeLa cells fixed with 4% formaldehyde (10 min) / permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

Flow Cyt

Lab

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labelling ERK1 with ab312910 at 1/50 dilution (1 ug) (Red) compared with a Rabbit monoclonal IgG (ab199091) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab303596 (right) or Rabbit IgG (monoclonal) Alexa Fluor® 488 (ab199091) isotype (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab303596) or Rabbit IgG (monoclonal) Alexa Fluor® 488 (ab199091) isotype (1x 106 in 100μl at 0.2 μg/ml (1/2500)) for 30 min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488 nm) and 525/40 bandpass filter. Events were gated on live single cells.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling ILT-4 with ab315183 at 1/500 dilution (0.1 ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control. Cells are co-stained with CD14 conjugated to Alexa Fluor® 647. Gated on viable cells.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab303573 (right) or Rabbit IgG (monoclonal) Alexa Fluor®488 (ab199091) isotype (left). Cells were fixed and permeabilized with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min on ice in 1x PBS containing 10 μg/ml human IgG and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab303573) or Rabbit IgG (monoclonal) Alexa Fluor®488 (ab199091) isotype (100 μl at 0.2 μg/ml (1/2500)) for 30 min on ice. The cells were simultaneously stained with CD4.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488 nm) and 525/40 bandpass filter.

Events were gated on live lymphocytes.

Flow Cyt

Lab

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Left). THP-1 (human monocytic leukemia monocyte, Right) cells labelling SIRP alpha with ab312902 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : Jurkat.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of permeabilized MDA-MB-231 (human breast adenocarcinoma epithelial cell, Left) / HepG2 (human hepatocellular carcinoma epithelial cell, Right) cells labelling ENPP1/PC1 with ab312900 at 1/50 dilution (1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab199091) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : MDA-MB-231.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / THP-1 (human monocytic leukemia monocyte, Right) cells labelling Iba1 with ab312913 at 1/5000 dilution (0.01 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab199091) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Negative control : MCF7 (PMID : 21509525).

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (human breast adenocarcinoma epithelial cell, Left) / HeLa (human cervix adenocarcinoma epithelial cell, Right) cells labeling CDKN2A/p14ARF with ab305363 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Negative control : MCF7 (PMID : 29904067).

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 293T (human embryonic kidney epithelial cell, Left) / Daudi (human Burkitt's lymphoma lymphoblast, Middle) / K562 (human chronic myelogenous leukemia lymphoblast, Right) cells labelling MICB with ab323890 at 1/50 dilution (1ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Gated on viable cells.
Negative control : 293T, Daudi (PMID : 28154561).

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized THP-1 (human monocytic leukemia monocyte, Left) and Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Right) cells labelling CD3D with ab198935 at 1/5000 dilution (0.01ug) / Red compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor^® 488) (ab199091) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Negative control : THP-1.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry scatter plots showing untreated human peripheral mononuclear cells (PBMCs) (top) and PBMCs activated with 5 µg/ml PHA for 18h (bottom), either stained with ab326135 (right) or isotype control (left). The cells were fixed and permeabilised using BD CytoFix/CytoPerm™ (20 min). The cells were incubated in 1x PBS containing 10 µg/ml human IgG, 10% normal goat serum and BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer (1/10) to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab326135) (1x 106 in 100µl at 1 µg/ml (1/500)) for 30min on ice. Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). The cells were simultaneously stained with CD3. Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing wild-type HeLa (green line) and GYS1 knockout HeLa stained with ab326126 (magenta line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326126) (1x 106 in 100µl at 1 µg/ml (1/500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HeLa WT cells (black line) and HeLa-GYS1 KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.This antibody gave a positive signal in wild-type HeLa fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling TLR8 with ab324390 at 1/50 dilution (1ug) / Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Cells were stained with anti-CD14 conjugated to BV421. Fixed with 2% PFA for 10 min followed by intracellularly staining with rabbit IgG or our our antibody.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing wild-type HCT-116 (green line) and HIF1a knockout HCT-116 stained with ab326123 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab326123) (1x 106 in 100µl at 1 µg/ml (1/500)) for 30min at 22°C.Isotype control antibody was Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (black line) in HCT-116 WT cells (black line) and HCT-116-HIF1a KO cells (grey line), used at the same concentration and conditions as the primary antibody. Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.This data was acquired using a Beckman Coulter CytoFlexLX flow cytometer.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype / 293T (human embryonic kidney epithelial cell) transfected with a TRDC (human TCR delta) protein expression vector containing a myc-His tag(Up Right) / 293T transfected with a TRGC1 protein (human TCR gamma) expression vector (Down Left) / 293T transfected with an empty vector containing a myc-His tag (Down Right) cells labelling TCR delta with ab317064 at 1/500 dilution (0.1ug)/ Up Right, Down Left and Down Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Flow Cyt

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Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Isotype (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with Myc-tagged ENPP3 overexpression construct (Middle) / 293T transfected with an empty vector containing a myc-His tag (Right) cells labelling ENPP3/B10 with ab319125 at 1/50 dilution (1ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody) (Blue).

Cells were co-stained with anti-myc conjugated to Alexa Fluor 647.
Gated on viable cells.

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a mouse CD27 expression vector (Middle) / 293T cells transfected with an empty expression vector (Right) cells labelling CD27 with ab324176 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Cells were co-stained with Myc tag conjugated to Alexa Fluor®647.

Flow Cyt

Supplier Data

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 2% paraformaldehyde fixed 0.1% Tween-20 permeabilized Isotype (Left) / 293T cells transfected with a mouse CCR4 expression vector containing a myc-His-tag® (Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Right) cells labelling CCR4 with ab324335 at 1/5000 dilution (0.01ug) / Middle and Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Cells were surface stained with isotype control or our antibody. Then fixed with 2% PFA for 10min followed by intracellularly stained with anti-myc tag conjugated to Alexa Fluor® 647.

Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab198978 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab198978, 1/500 dilution) for 30 min at 22°C.

Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor^® 488 (ab199091) used at the same concentration and conditions as the primary antibody. Unlabeled sample (blue line) was also used as a control.

Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometry overlay histogram showing left HeLa positive cells and right negative HeLa VCL KO stained with ab196454 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab196454) (1x 10⁶ in 100μl at 0.008μg/ml (1/62500)) for 30min at 22°C.

Isotype control antibody (black line) was Recombinant Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Flow Cyt

Supplier Data

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling Survivin with ab307852 at 1/50 dilution (1.0ug) (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / HEK-293T (human embryonic kidney epithelial cell) transfected with myc-tagged IL-22 expression vector containing a myc tag (Right) cells labelling IL-22 with ab317658 at 1/500 dilution (0.1ug) / Right (Red) compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Flow Cyt

Supplier Data

Flow Cytometry - Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (AB199091)

Flow cytometric analysis of Human PBMC(human peripheral blood mononuclear cell) cells labelling CD45 with ab315958 at 1/5000 dilution (0.01ug)/Right compared with a Rabbit IgG monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488) (ab199091) / Left isotype control.

Human PBMCs are co-stained with CD3 conjugated Alexa Fluor®647.Gated on viable cell.

不同偶联物与剂型 (15)

Unconjugated

Rabbit IgG, monoclonal [EPR25A] - Isotype Control
Carrier free

Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free
Carrier free

Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free)
660 APC

APC Rabbit IgG, monoclonal [EPR25A] - Isotype Control
421 Alexa Fluor® 405

Alexa Fluor® 405 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
565 Alexa Fluor® 555

Alexa Fluor® 555 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
603 Alexa Fluor® 568

Alexa Fluor® 568 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
617 Alexa Fluor® 594

Alexa Fluor® 594 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
665 Alexa Fluor® 647

Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
775 Alexa Fluor® 750

Alexa Fluor® 750 Rabbit IgG, monoclonal [EPR25A] - Isotype Control
Biotin

Biotin Isotype Control [EPR25A]
519 FITC

FITC Rabbit IgG, monoclonal [EPR25A] - Isotype Control
HRP

HRP Rabbit IgG, monoclonal [EPR25A] - Isotype Control
578 PE

PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control
675 PerCP

PerCP Rabbit IgG, monoclonal [EPR25A] - Isotype Control

关键信息

宿主种属

Rabbit

克隆

Monoclonal

克隆号

EPR25A

亚型

IgG

偶联物

Alexa Fluor® 488

激发波长/发射波长

Ex: 495nm, Em: 519nm

不含载体蛋白

反应种属

Human

应用

Flow Cyt (Intra), Flow Cyt, IHC-P, ICC/IF

applications

特异性

Please note Abcam have optimised the validation of this product. In our hands, we observe an increase in background signal intensity with the use of Triton X-100 and would recommend using an alternative permeabilisation method such as methanol or saponin.

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"}, "IHCP" : {"fullname" : "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)", "shortname":"IHC-P"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCyt-species-checked": "guaranteed", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "guaranteed", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" }, "": { "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/500", "ICCIF-species-notes": "", "FlowCyt-species-checked": "testedAndGuaranteed", "FlowCyt-species-dilution-info": "1/500", "FlowCyt-species-notes": "Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.", "FlowCytIntra-species-checked": "", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "", "IHCP-species-checked": "", "IHCP-species-dilution-info": "", "IHCP-species-notes": "" } } }

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产品详情

Product Specifications
Alexa Fluor^® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199091) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt, Flow Cyt (Intra) and ICC/IF.

Quality and Validation
Alexa Fluor^® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199091) has been cited over 25 times in peer reviewed journals and is trusted by the scientific community.
Flourescent conjugated antibodies are ideal tools for multiplex IHC and flow cytometry experiments.
Alexa Fluor^® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199091) has 25 independent reviews from customers.
Alexa Fluor^® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199091) specifically detects rabbit IgG and is sold in 100 µL selling sizes.

Related Products
Conjugation-ready, carrier free format available for antibody clone (EPR25A) - ab210849.

Immunogen
Chemical / Small Molecule conjugated to keyhole limpet haemocyanin. KLH is a copper containing oxygen carrier occurring freely dissolved in the hemolymph of many molluscs and arthropods. KLH forms a large complex composed of ~50 kDa subunits.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

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性能和储存信息

形式

Liquid

纯化工艺

Affinity purification Protein A

存储溶液

pH: 7.4 Preservative: 0.02% Sodium azide Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA

运输条件

Blue Ice

分装信息

Upon delivery aliquot

储存信息

Avoid freeze / thaw cycle|Store in the dark

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产品实验方案

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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靶点信息

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文献 (33)

Recent publications for all applications. Explore the full list and refine your search

The Journal of clinical investigation 135: PubMed40100289

2025

AMPK is necessary for Treg functional adaptation to microenvironmental stress during malignancy and viral pneumonia.

Applications

Unspecified application

Species

Unspecified reactive species

Manuel A Torres Acosta,Jonathan K Gurkan,Qianli Liu,Nurbek Mambetsariev,Carla Reyes Flores,Kathryn A Helmin,Anthony M Joudi,Luisa Morales-Nebreda,Kathleen Cheng,Hiam Abdala-Valencia,Samuel E Weinberg,Benjamin D Singer

Heliyon 10:e40734 PubMed39717606

2024

Serum ADAM17 levels pre-antiviral therapy correlate with HIV patient immune reconstitution.

Applications

Unspecified application

Species

Unspecified reactive species

Ying Lin,Ming Chen,Xuelian Zhu, Xiaoya Cui,Suyu Sun,Xingzhong Hu

Cell reports. Medicine 5:101869 PubMed39657666

2024

Miltefosine reinvigorates exhausted T cells by targeting their bioenergetic state.

Applications

Unspecified application

Species

Unspecified reactive species

Xingying Zhang,Chenze Zhang,Shan Lu,Jingxi Dong,Na Tang,Yao Wang,Weidong Han,Xi Pan,Xiang Zhang,Duan Liu,Ng Shyh-Chang,Yu Wang,Guihai Feng,Haoyi Wang

Journal of orthopaedic translation 45:211-225 PubMed38586591

2024

IRF1 governs the expression of SMARCC1 via the GCN5-SETD2 axis and actively engages in the advancement of osteoarthritis.

Applications

Unspecified application

Species

Unspecified reactive species

Dong Wang,Yujun Zhang,Liangping Zhang,Du He,Lan Zhao,Zhimin Miao,Wei Cheng,Chengyue Zhu,Li Zhu,Wei Zhang,Hongting Jin,Hang Zhu,Hao Pan

Journal of translational medicine 22:5 PubMed38169393

2024

PIGT promotes cell growth, glycolysis, and metastasis in bladder cancer by modulating GLUT1 glycosylation and membrane trafficking.

Applications

Unspecified application

Species

Unspecified reactive species

Mingyue Tan,Qi Pan,Chao Yu,Xinyu Zhai,Jianyi Gu,Le Tao,Dongliang Xu

Journal of experimental & clinical cancer research : CR 42:304 PubMed37974213

2023

Targeting FAcilitates Chromatin Transcription complex inhibits pleural mesothelioma and enhances immunotherapy.

Applications

Unspecified application

Species

Unspecified reactive species

Anand Singh,Nathanael Pruett,Shivani Dixit,Sudheer K Gara,Haitao Wang,Roma Pahwa,David S Schrump,Chuong D Hoang

Cellular & molecular biology letters 28:91 PubMed37946128

2023

Irf7 regulates the expression of Srg3 and ferroptosis axis aggravated sepsis-induced acute lung injury.

Applications

Unspecified application

Species

Unspecified reactive species

Xinyu Ling,Shiyou Wei,Dandan Ling,Siqi Cao,Rui Chang,Qiuyun Wang,Zhize Yuan

Small (Weinheim an der Bergstrasse, Germany) 19:e2300578 PubMed37423970

2023

A Biomimetic Peptide Functions as Specific Extracellular Matrix for Quiescence of Stem Cells against Intervertebral Disc Degeneration.

Applications

Unspecified application

Species

Unspecified reactive species

Yuan Li,Kuo Zhang,Xuezheng Ai,Qingshi Zhang,Lu Jiang,Jiang Long,Huange Xu,Chencheng Feng,Yaqing Zhang,Guoke Tang,Fanli Chong,Lei Wang,Bo Huang

Molecular cancer therapeutics : PubMed37196156

2023

An oncolytic poxvirus encoding hNIS, shows anti-tumor efficacy and allows tumor imaging in a liver cancer model.

Applications

Unspecified application

Species

Unspecified reactive species

Shyambabu Chaurasiya,Hannah Valencia,Zhifang Zhang,Sang-In Kim,Annie Yang,Jianming Lu,Yanghee Woo,Susanne G Warner,Nicholas J Ede,Yuman Fong

Biomolecules 13: PubMed37189383

2023

Engineering of a Bispecific Nanofitin with Immune Checkpoint Inhibitory Activity Conditioned by the Cross-Arm Binding to EGFR and PDL1.

Applications

Unspecified application

Species

Unspecified reactive species

Perrine Jacquot,Javier Muñoz-Garcia,Maurine Fleury,Denis Cochonneau,Rémi Gaussin,Elise Enouf,Caroline Roze,Emilie Ollivier,Mathieu Cinier,Dominique Heymann

View all publications

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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。

详情请参阅我们的条款与条件。

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Alexa Fluor® 488 Rabbit IgG, monoclonal [EPR25A] - Isotype Control