Anti p53 monoclonal antibody do 1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

The Anti-p53 monoclonal antibody (DO-1) is a laboratory reagent produced by Santa Cruz Biotechnology. It is designed to detect the p53 protein, a key tumor suppressor involved in cell cycle regulation and apoptosis. The antibody can be used in various immunological techniques, such as Western blotting, immunoprecipitation, and immunohistochemistry, to identify and study the p53 protein in biological samples.

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Lab products found in correlation

Immobilon p membrane, by Merck Group (1 mentions) Anti max antibody, by Santa Cruz Biotechnology (1 mentions) Anti myc n 262, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

Anti-p53 (329 citations) P53 (DO-1, (97 citations) Anti-p53 (DO-1, (81 citations) Anti-p53 antibody (41 citations) Anti-p53 antibody (DO-1) (24 citations) P53 antibody (16 citations) P53 antibody DO-1 (13 citations) Mouse monoclonal anti-p53 DO-1 antibody (2 citations)

3 protocols using anti p53 monoclonal antibody do 1

Protein Extraction and Western Blot Analysis

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Cell aliquots (5×10⁶) were removed at various time points, collected by centrifugation and washed once in PBS, then lysed in extraction buffer (20 mM Tris-HCl at pH 7.4, 150 mM NaCl, 1% Triton X-100, 5 mM EDTA, 1 mM Na₃VO₄, 10 mM NaF, 10 mM Na₄P₂O₇, 1 mM PMSF, 1 µg/mL leupeptin, and 0.1 U/mL of aprotinin) and cleared by centrifugation for 10 minutes at 4°C to obtain whole-cell extracts. Each 50 µg sample of total protein was loaded onto 4%–15% gradient sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels and blotted onto poly(vinylidene) difluoride Immobilon-P membranes (EMD Millipore, Billerica, MA, USA).
The following antibodies were used: anti-p53 monoclonal antibody (DO-1) (Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-Ran antibody (SCB), anti-p21^waf1/cip1 antibody (Ab-1) (Oncogene Science, Cambridge, MA, USA), anti-phospho p53 (Ser15) mouse monoclonal antibody (Cell Signaling Technology, Danvers, MA, USA), and anti-phospho p53 (Ser20) rabbit polyclonal antibody (CST).

Bainer R.O., Trendowski M.R., Cheng C., Pei D., Yang W., Paugh S.W., Goss K.H., Skol A.D., Pavlidis P., Pui C.H., Gilliam T.C., Evans W.E, & Onel K. (2017). A p53-regulated apoptotic gene signature predicts treatment response and outcome in pediatric acute lymphoblastic leukemia. Cancer Management and Research, 9, 397-410.

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Immunoprecipitation and Western Blot Analysis of p53, MYCN, and MYC

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Crude nuclear protein extract from Nutlin-3a-treated (10 μM for 4 h) IMR-32 cells and Hela cells were prepared. Nuclei were isolated using hypotonic NP-40 buffer (10 mM HEPES, 50 mM NaCl, 1 mM DTT, 1 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1 mM sodium fluoride, 1 mM PMSF. cOmplete Protease Inhibitor Cocktail, and 0.2% NP-40) and lysed via sonication in nuclei lysis buffer (50 mM Tris HCl pH 8.00, 150 mM NaCl, 1 mM DTT, 1 mM sodium pyrophosphate, 1 mM sodium orthovanadate, 1 mM sodium fluoride, 1 mM PMSF, cOmplete Protease Inhibitor Cocktail, and 0.05% NP-40). One milligram of crude nuclear protein extract was incubated overnight with 2 μg of anti-p53 antibody (Ab-7, Calbiochem) or 2 μg of control sheep IgG at 4°C. Recombinant Protein G agarose resin (Invitrogen) was used to purify protein complexes. Following three washes with 0.15% NP-40 in nuclei lysis buffer, protein samples were eluted by boiling resin in Laemmli sample buffer, separated using SDS-PAGE, and analyzed using Western blot with an anti-MYCN monoclonal antibody (B8.4.B, Santa Cruz Biotechnology), anti-p53 monoclonal antibody (DO-1, Santa Cruz Biotechnology), anti-MAX antibody (C17, Santa Cruz Biotechnology), and anti-MYC (N262, Santa Cruz Biotechnology).

Agarwal S., Milazzo G., Rajapakshe K., Bernardi R., Chen Z., Barberi E., Koster J., Perini G., Coarfa C, & Shohet J.M. (2018). MYCN acts as a direct co-regulator of p53 in MYCN amplified neuroblastoma. Oncotarget, 9(29), 20323-20338.

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Vero Cell Apoptosis Analysis

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Vero cells were treated with or without BSP-II for 24 h. Positive control cells were treated with Dox. Cell proteins were collected and subjected to Western blotting as previously described [14 (link)] with anti-p53 monoclonal antibody (DO-1; Santa Cruz Biotechnology, USA), anti-Bax polyclonal antibody (N-20; Santa Cruz Biotechnology), and anti-β-actin monoclonal antibody (AC-15; Sigma).

Zhou G.F., Liu Q.T., Zhou B., Qiu Y.F., Liu X.D., Ma Z.Y., Feng X.L., Cao R.B, & Chen P.Y. (2015). The potential molecular effects of bursal septpeptide II on immune induction and antitumor activity. Journal of Veterinary Science, 16(3), 325-331.

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