重组Anti-mCherry抗体[EPR20579]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mCherry antibody [EPR20579] (AB213511)

Immunohistochemistry analysis of (A) 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry. (B) 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty expression vector labeling mCherry with ab213511 at 1/100 dilution followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as the secondary antibody. Sections were fixed with paraffin. Hematoxylin was used as the couterstain. Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on 293T cell pellets transfected with a His-tagged mCherry, no staining on the 293T cell pellets transfected with an empty expression vector. The section was incubated with ab213511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• Flow Cyt

Lab

Flow Cytometry - Anti-mCherry antibody [EPR20579] (AB213511)

Flow cytometry analysis of 293T (human embryonic kidney epithelial cell) labeling mCherry with ab213511 at 1/500 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/5000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% methanol. Isotype control : Rabbit monoclonal IgG (ab172730) (Left).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-mCherry antibody [EPR20579] (AB213511)

Immunocytochemistry/ Immunofluorescence analysis of 293T (human embryonic kidney epithelial cell) labeling mCherry with ab213511 at 1/100 dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. DAPI was used as nuclear counterstain. Confocal image showing positive staining in 293T cells transfected with a mCherry expression vector containing a myc tag. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mCherry antibody [EPR20579] (AB213511)

Immunohistochemistry analysis of paraffin embedded human tonsil tissue sections labeling mCherry with ab213511 at 1/100 dilution followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as the secondary antibody. Sections were fixed with paraffin. Hematoxylin was used as the couterstain. Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. No staining on human tonsil. The section was incubated with ab213511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mCherry antibody [EPR20579] (AB213511)

Immunohistochemistry analysis of paraffin embedded rat spleen tissue sections labeling mCherry with ab213511 at 1/100 dilution followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as the secondary antibody. Sections were fixed with paraffin. Hematoxylin was used as the couterstain. Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. No staining on rat spleen. The section was incubated with ab213511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mCherry antibody [EPR20579] (AB213511)

Immunohistochemistry analysis of paraffin embedded mouse spleen tissue sections labeling mCherry with ab213511 at 1/100 dilution followed by LeicaDS9800 (Bond™ Polymer Refine Detection) as the secondary antibody. Sections were fixed with paraffin. Hematoxylin was used as the couterstain. Antigen retrieval was heat mediated with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. No staining on mouse spleen. The section was incubated with ab213511 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-mCherry antibody [EPR20579] (AB213511)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labelling mCherry with ab213511 at 1/100 dilution

ab213511 showed no staining on liver tissue from mCherry-EGFP-LC3 fusion protein knock-in mice, which may be due to epitope masking by LC3 or EGFP.

The section was incubated with ab213511 or ab125096 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

The tissue samples were kindly provided by GemPharmatech

• I-ELISA

Lab

Indirect ELISA - Anti-mCherry antibody [EPR20579] (AB213511)

ELISA analysis of mPlum fluorescent protein,Osred fluorescent protein,tdTomato fluorescent protein,mcherry fluorescent protein at 1000 ng/mL with ab213511 at 1000-0 ng/mL. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-mCherry antibody [EPR20579] (AB213511)

mCherry was immunoprecipitated from 0.35 mg of 293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry whole cell lysate with ab213511 at 1/20 dilution. Western blot was performed from the immunoprecipitate using ab213511 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : 293T transfected with a His-tagged mCherry whole cell lysate 10 μg (Input).

Lane 2 : ab213511 IP in 293T transfected with a His-tagged mCherry whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab213511 in 293T transfected with a His-tagged mCherry whole cell lysate.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-mCherry antibody [EPR20579] (ab213511)

false

• WB

Supplier Data

Western blot - Anti-mCherry antibody [EPR20579] (AB213511)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-mCherry antibody [EPR20579] (ab213511) at 1/1000 dilution

Lane 1:

293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with an empty expression vector whole cell lysate at 10 µg

Lane 2:

293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged mCherry whole cell lysate at 10 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

Predicted band size: 26 kDa

Observed band size: 29 kDa

true

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-mCherry antibody [EPR20579] (AB213511)

Blocking/Dilution buffer : 5% NFDM/TBST.

Green fluorescent protein (GFP) contains aa1-238 with His-tag.

All lanes:

Western blot - Anti-mCherry antibody [EPR20579] (ab213511) at 1/1000 dilution

All lanes:

Green fluorescent protein (GFP) at 0.01 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

Predicted band size: 26 kDa

true

Exposure time: 3min