txt: This is a knockout-validated antibody summary, based on the publication "Suppression of KRas-mutant cancer through the combined inhibition of KRAS with PLK1 and ROCK", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western blot of p21 in HCT-116 (KRAS mutant, p21 wild type) and HCT-116 (KRAS mutant, p21−/−). β-actin was used as loading control. From [1].

Rabbit monoclonal IgG

Company: Abcam

Antibody: p21

Catalog number: ab109520

Summary: Rabbit monoclonal IgG against a synthetic peptide within human p21 aa 1-100. Reacts with human protein. Suitable for western blot, immunoprecipitation, immunocytochemistry/immunofluorescence, immunohistochemistry (paraffin) and flow cytometry.

Western blot

HCT-116 (KRAS mutant, p21 wild type) and HCT-116 (KRAS mutant, p21−/−) cells. Whole-cell or tumour tissue extracts were prepared by radioimmunoprecipitation assay buffer (150 mM sodium chloride, 50 mM Tris pH 8.0, 1% Triton X-100, 0.5% sodium deoxycholate, 0.1% SDS) supplemented with 10 mM NaF, 1 mM Na3VO4, 5 mM EDTA, 1 mM EGTA, 5 μg ml−1 leupeptin, 1 μg ml−1 pepstatin A, 1 mM phenylmethylsulfonyl fluoride, and protease and phosphatase inhibitor cocktail (Calbiochem).

5% bovine serum albumin for 1 h at room temperature.

1:2,000 dilution in Primary Antibody Dilution Buffer (Beyotime Biotechnology, China) at 4 °C overnight.

1:10,000 diluted IRDye 800CW Goat anti-Rabbit IgG (926–32211, LI-COR Biosciences).

Odyssey Infrared Imaging System (Odyssey, LI-COR).

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.