Recombinant Anti-PCNA antibody [EPR3821] (ab92552)

Western blotting PCNA

Experiment
Western blotting PCNA
Product
Recombinant Anti-PCNA antibody [EPR3821] (ab92552) from Abcam
Manufacturer
Abcam

Protocol tips

Upstream tips
-These buffers may be stored at 4°C for several weeks or aliquoted and stored at -20°C for up to a year.
Protocol tips
-Rabbit (1:5000)
-To reduce and denature your samples, boil each cell lysate in sample buffer at 100°C for
5 min. Lysates can be aliquoted and stored at -20°C for future use.

Publication protocol

Cultured or transfected cells were harvested and washed twice with PBS. Then, the cells were lysed in ice-cold radio immunoprecipitation assay buffer (Beyotime, Shanghai, People’s Republic of China) containing 0.01% protease and phosphatase inhibitor (Sigma, Shanghai, People’s Republic of China) and incubated on ice for 30 min. Cell lysis was centrifuged at 12,000× g for 10 min at 4°C, and the proteins in supernatant were obtained (20−30 μg) and separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis gel, then transferred electrophoretically to a polyvinylidene fluoride membrane (Millipore, Shanghai, People’s Republic of China). The membrane was blocked with 5% bovine serum albumin in tris-buffered saline (TBS) and Polysorbate 20 (also known as Tween 20) and incubated with primary antibodies against ARHGAP1 (Ab194425, 1:1,000 dilution; Abcam), matrix metallopeptidase 2 (MMP2; Ab92536, 1:1,000 dilution; Abcam), zinc finger E-box binding homeobox 1 (ZEB1; Ab124512, 1:1,000 dilution; Abcam), Cyclin B1 (CCNB1; #12231, 1:1,000 dilution; Cell Signaling Technology, Inc.), twist family bHLH transcription factor 1 (Twist; Ab175430, 1:500 dilution; Abcam), proliferating cell nuclear antigen (PCNA; Ab92552, 1:5,000 dilution; Abcam) and GAPDH (#5174, 1:1,500 dilution; CST). Blots were then incubated for 1 h at 37°C with goat anti-mouse or anti-rabbit secondary antibody (Beyotime), and the intensities were measured using enhanced chemiluminescence (Thermo Fisher Scientific Inc.)

Full paper   Login or join for free to view the full paper.

Reviews

Recombinant Anti-PCNA antibody [EPR3821] (ab92552) from Abcam has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

Discussion

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting PCNA using Recombinant Anti-PCNA antibody [EPR3821] (ab92552) from Abcam.

View full paper   Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Abcam for Recombinant Anti-PCNA antibody [EPR3821] (ab92552) below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting PCNA using Recombinant Anti-PCNA antibody [EPR3821] (ab92552) from Abcam. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms