Anti-mCherry antibody [EPR20579]

• 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).

• 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

Lab

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

Blocking and diluting buffer and concentration : 5% NFDM /TBST.
DYRK1 has been shown to undergo degradation (PMID : 27807027)

Lanes 1 - 3:

Western blot - Anti-DYRK1A + DYRK1B antibody [EPR24132-61] (<a href='/en-us/products/primary-antibodies/dyrk1a-antibody-epr24132-61-ab259869'>ab259869</a>) at 1/1000 dilution

Lanes 1 - 3:

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

Lane 1:

293T cells transfected with an empty vector containing a mCherry tag whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a full length human DYRK1A expression vector containing a mCherry tag whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a full length human DYRK1B expression vector containing a mCherry tag whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 86 kDa

false

Exposure time: 3s

• 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.

• 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='/en-us/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

• 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='/en-us/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