p21 Antibody (F-5): sc-6246

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Protocol tips
-Mouse (1:1000) -Use clean forceps to gently handle the blot from the corner without creasing the membrane. Do not write on the blot with pen or marker, as the ink can fluoresce and cause background.

Publication protocol

Proteins were extracted by boiling cell pellets in sample buffer (0.35 M Tris pH 6.8, 0.1 g/ml sodium dodecyl sulphate, 93 mg/ml dithiothreitol, 30% glycerol, 50 μg/ml bromophenol blue), resolved by SDS-PAGE, then electroblotted onto Immobilon-P membranes (Merck Millipore). Following blocking in 5% dried skimmed milk (Marvel) dissolved in TBS-T (50 mM Tris pH 7.6, 150 mM NaCl, 0.1% Tween-20), membranes were incubated with primary antibodies (BUB3 1:1000; HUS1 1:500; LAMIN B1 1:1000; PARG 1:1000; c-MYC 1:3500; TP53 1:1000; P21 1:100; RFC2 1:1000; RECQL1 1:1000; RPA32 1:1000; p-RPA32 1:500; TAO1 1:1000; TIMELESS 1:1000) overnight at 4°C. Membranes were then washed three times in TBS-T and incubated for at least 1 hr with appropriate horseradish-peroxidase-conjugated secondary antibodies (1:2000). After washing in TBS-T, bound secondary antibodies were detected using either EZ-Chemiluminescence Reagent (Geneflow Ltd) or Luminata™ Forte Western HRP Substrate (Merck Millipore) and a Biospectrum 500 imaging system (UVP) or ChemiDoc™ Touch Imaging System (BioRad).

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

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Western blotting p21 using p21 Antibody (F-5): sc-6246 from Santa Cruz Biotechnology.

Manufacturer protocol

Download the product protocol from Santa Cruz Biotechnology for p21 Antibody (F-5): sc-6246 below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing Western blotting p21 using p21 Antibody (F-5): sc-6246 from Santa Cruz Biotechnology. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.