Anti-CD19 antibody [RM2072]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM2072] (AB322345)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD19 with ab322345 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human tonsil.
The section was incubated with ab322345 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD3 conjugated to Brilliant Violet 421.
Gated on viable cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [RM2072] (AB322345)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling CD19 with ab322345 at 1/50 (9.78 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing membranous staining in Raji cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : Jurkat (PMID : 19147785).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM2072] (AB322345)

Immunohistochemical analysis of paraffin-embedded Human non-Hodgkin's lymphoma tissue labeling CD19 with ab322345 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human non-Hodgkin's lymphoma.
The section was incubated with ab322345 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of Raji (human Burkitt's lymphoma B lymphocyte) (Right) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood) (Left) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM2072] (AB322345)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling CD19 with ab322345 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in human cerebrum.
The section was incubated with ab322345 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD20 conjugated to Alexa Fluor®647.
Gated on viable cells.

• IP

Supplier Data

Immunoprecipitation - Anti-CD19 antibody [RM2072] (AB322345)

CD19 was immunoprecipitated from 0.35 mg Raji whole cell lysate with ab322345 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322345 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : ab322345 IP in Raji whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322345 in Raji whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-CD19 antibody [RM2072] (ab322345) at 1/30 dilution

All lanes:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 81s

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of Mouse PBMC (mouse peripheral blood mononuclear cell) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD45R/B220 conjugated to Alexa Fluor®647 (660/20BP).
Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM2072] (AB322345)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD19 with ab322345 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in mouse spleen.
The section was incubated with ab322345 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [RM2072] (AB322345)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized WEHI-231 (mouse B cell lymphoma B lymphocyte) cells labelling CD19 with ab322345 at 1/50 (9.78 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in WEHI-231 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Negative control : CTLL-2 (PMID : 19147785).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of Mouse PBMC (mouse peripheral blood mononuclear cell) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti human CD3 conjugated to Brilliant Violet 421.
Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD19 antibody [RM2072] (AB322345)

Flow cytometric analysis of WEHI-231 (mouse B cell lymphoma B lymphocyte) (Right) / CTLL-2 (mouse T lymphocyte) (Left) cells labelling CD19 with ab322345 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM2072] (AB322345)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD19 with ab322345 at 1/100 (4.89 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in mouse cerebrum.
The section was incubated with ab322345 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IP

Supplier Data

Immunoprecipitation - Anti-CD19 antibody [RM2072] (AB322345)

CD19 was immunoprecipitated from 0.35 mg WEHI-231 whole cell lysate with ab322345 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab322345 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate
Lane 2 : ab322345 IP in WEHI-231 whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab322345 in WEHI-231 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-CD19 antibody [RM2072] (ab322345) at 1/30 dilution

All lanes:

WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD19 antibody [RM2072] (AB322345)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : Human skeletal muscle, mouse heart.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD19 antibody [RM2072] (ab322345) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate at 20 µg

Lane 2:

Human spleen tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse lymph node tissue lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60-120 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-CD19 antibody [RM2072] (AB322345)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

CD19 is a glycoprotein of approximately 100-120 kDa and detected as a 60-75 kDa band after treated with Protein Deglycosylation MIX II.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD19 antibody [RM2072] (ab322345) at 1/1000 dilution

Lane 1:

Untreated Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Raji cell lysate treated with Protein Deglycosylation Mix II at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60-120 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CD19 antibody [RM2072] (AB322345)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Negative control : CTLL-2 and Jurkat (PMID : 19147785).

The molecular weight observed is consistent with what has been described in the literature (PMID : 26516065; PMID : 14530327)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD19 antibody [RM2072] (ab322345) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

Ramos (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

NAMALWA (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 6:

A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 7:

WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 8:

CTLL-2 (mouse T lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 60-120 kDa,36 kDa

false

Exposure time: 180s