Anti-PUMA antibody [EPR26854-239]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PUMA antibody [EPR26854-239] (AB324435)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 (human prostate adenocarcinoma epithelial cell, Left) / LoVo (human colorectal adenocarcinoma epithelial cell, Right) cells labelling PUMA with ab324435 at 1/500 dilution (0.1ug) (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.

Low expression : PC-3.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PUMA antibody [EPR26854-239] (AB324435)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized LoVo (human colorectal adenocarcinoma epithelial cell) cells labelling PUMA with ab324435 at 1/50 (9.76 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/ab150081 1000 2ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in LoVo cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : PC-3.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/ab7291 1000 1ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324435 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-PUMA antibody [EPR26854-239] (AB324435)

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

The expression of PUMA is upregulated in response to doxorubicin treatment.

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

All lanes:

Western blot - Anti-PUMA antibody [EPR26854-239] (ab324435) at 1/1000 dilution

Lane 1:

Untreated A549 (human lung carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

A549 treated with 1µM doxorubicin for 48 hours whole cell lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 25 kDa,36 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-PUMA antibody [EPR26854-239] (AB324435)

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

Low expression : PC-3, AsPC-1.

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

All lanes:

Western blot - Anti-PUMA antibody [EPR26854-239] (ab324435) at 1/1000 dilution

Lane 1:

22Rv1 (human prostate carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

AsPC-1 (human Pancreas epithelial cell) whole cell lysate at 50 µg

Secondary

All lanes:

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

Observed band size: 25 kDa,36 kDa

false

Exposure time: 59s