p21/CIP1/CDKN1A Antibody (WA-1)

Immunohistochemistry-Paraffin: p21/CIP1/CDKN1A Antibody (WA-1) [NBP2-29463] - Formalin-fixed, paraffin-embedded human bladder carcinoma stained with p21 Monoclonal Antibody (WA-1).

• Reactivity: Hu, Mu, Rt, Pm, Pm
• Applications: WB, Flow, ICC/IF, IHC, IHC-P
• Clone: WA-1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

p21/CIP1/CDKN1A Antibody (WA-1) Summary

• Immunogen: Human recombinant p21/CIP1/CDKN1A protein
• Localization: Nuclear
• Specificity: This MAb recognizes a 21kDa protein, identified as the p21WAF1 tumor suppressor protein. This MAb is highly specific to p21 and shows no cross-reaction with other closely related mitotic inhibitors. p21WAF1 is a specific inhibitor of cdk s and a tumor suppressor involved in the pathogenesis of a variety of malignancies. The expression of this gene acts as an inhibitor of the cell cycle during G1 phase and is tightly controlled by the tumor suppressor protein p53. Its expression is induced by the wild type, but not mutant, p53 suppressor protein. Normal cells generally display a rather intense nuclear p21 expression. Loss of p21 expression has been reported in many carcinomas (gastric carcinoma, non-small cell lung carcinoma, thyroid carcinoma). In ELISA, MAb WA-1 is useful either as a solid phase or for detection of p21 protein.
• Isotype: IgG1 Kappa
• Clonality: Monoclonal
• Host: Mouse
• Gene: CDKN1A
• Purity: Protein A or G purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 0.5-1ug/million cells
  • Immunocytochemistry/Immunofluorescence 1-2ug/ml
  • Immunohistochemistry 0.5-1ug/ml
  • Immunohistochemistry-Paraffin 0.5-1ug/ml
  • Western Blot
• Application Notes: Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
  Optimal dilution for a specific application should be determined.
  Use in Western blot reported in scientific literature (PMID 28137862).
• Theoretical MW: 21 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Packaging, Storage & Formulations

• Storage: Store at 4C.
• Buffer: 10 mM PBS with 0.05% BSA
• Preservative: 0.05% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Protein A or G purified

Notes

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml (NBP2-33084).

Alternate Names for p21/CIP1/CDKN1A Antibody (WA-1)

• CAP20cyclin-dependent kinase inhibitor 1
• CDK-interacting protein 1
• CDKN1A
• CDKN1melanoma differentiation associated protein 6
• CIP1
• CIP1WAF1CDK-interaction protein 1
• cyclin-dependent kinase inhibitor 1A (p21, Cip1)
• MDA6
• MDA-6
• Melanoma differentiation-associated protein 6
• p21
• p21CIP1
• p21Cip1/Waf1
• PIC1
• SDI1DNA synthesis inhibitor
• wild-type p53-activated fragment 1

Background

p21 (WAF1), a member of the cycle dependent kinase (CDK) inhibitor family, was first identified as a tumor suppressor or anti-oncogenic protein and negative regulator of the cell cycle. Over the years it became apparent that p21 has broad functions in regulating fundamental cellular programs including proliferation, differentiation, migration, senescence, and has both anti-oncogenic and oncogenic properties (reviewed in Romanov, 2012). Many antibody and other studies have demonstrated that p21 levels are frequently elevated in situations associated with reduced proliferation, including differentiation and senescence, and decreased in proliferative states. Subcellular localization may significant and nuclear antibody staining may be indicative of p21 functioning as a cell cycle inhibitor, tumor suppressor, or in a senescence program. In contrast, cytoplasmic antibody staining may indicate p21 is acting an oncogene through regulatation of migration, proliferation, or apoptosis.

It is interesting to note that p21 expression is induced by wild type, but not mutant p53. The HJ21/WA-1 antibody clone has been used to demonstrate this phenomenon by western blot (Blaydes, 2001). The inability of mutant p53 to induce p21 essentially means that normal functions of p21 are compromised when p53 gene mutations are present. Since p53 gene mutations are present in up to 50% of human cancers, the loss of p21 function in cancer is significant.

In its normal functioning state, p21 binds to cyclin/CDK complexes. When it binds to these complexes, it inhibits their kinase activity which, in turn, stops cell cycle progression and hence p21 gains its reputation as a mitotic cell cycle inhibitor. In this regard, the antibody showed that decreased Cdk2-cyclin E1 activity corresponded with a decrease in cyclin E1 and increase in p21 protein levels (White, 2005).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for p21/CIP1/CDKN1A Antibody (NBP2-29463)(15)

Publications using NBP2-29463

• Saleh T, Alhesa A, Al-Balas M et al. Expression of therapy-induced senescence markers in breast cancer samples upon incomplete response to neoadjuvant chemotherapy Bioscience reports May 28 2021 [PMID: 33948615] (IHC, Human)
• Kaczorowski A, Tolstov Y, Falkenstein M et al. Rearranged ERG confers robustness to prostate cancer cells by subverting the function of p53 Urol. Oncol. Jul 13 2020 [PMID: 32674955] (WB, Human)
• Couture R, Mora N, Al Bittar S, et al. Luteolin modulates gene expression related to steroidogenesis, apoptosis, and stress response in rat LC540 tumor Leydig cells Cell Biol. Toxicol. 2019 Jun 14 [PMID: 31201582]
• Aratani S, Tagawa M, Nagasaka S et al. Radiation-induced premature cellular senescence involved in glomerular diseases in rats. Sci Rep. Nov 14 2018 [PMID: 30429495] (IHC-P, Rat)
• Huang X, Huang R, Gou S et al. Platinum(IV) complexes conjugated with phenstatin analogue as inhibitors of microtubule polymerization and reverser of multidrug resistance Bioorg. Med. Chem. 2017 Jul 08 [PMID: 28728896] (Human)
• Xiang J, Yang S, Xin N et al. DYRK1A regulates Hap1-Dcaf7/WDR68 binding with implication for delayed growth in Down syndrome Proc. Natl. Acad. Sci. U.S.A Jan 30 2017 [PMID: 28137862] (WB, Mouse)
• Ge X, Pettan-Brewer C, Morton J et al. Mitochondrial catalase suppresses naturally occurring lung cancer in old mice. Pathobiol Aging Age Relat Dis 2015 Sep 24 [PMID: 26400209] (IHC, Mouse)
• Romanov VS, Pospelov VA, Pospelova TV. Cyclin-dependent kinase inhibitor p21(Waf1): contemporary view on its role in senescence and oncogenesis. Biochemistry (Mosc). 2012 Jun [PMID: 22817456]
• Koike M, Yutoku Y, Koike A. Accumulation of p21 proteins at DNA damage sites independent of p53 and core NHEJ factors following irradiation. Biochem Biophys Res Commun. 2011 Aug 19 [PMID: 21787748]
• Nobre RJ, Herraez-Hernandez E, Fei JW et al. E7 oncoprotein of novel human papillomavirus type 108 lacking the E6 gene induces dysplasia in organotypic keratinocyte cultures. J Virol. 2009 Apr [PMID: 19153227] (WB, IHC-P, Human)
  
  Details:
  Human keratinocyte oragnotypic raft cultures: IHC (paraffin), Fig3A; WB, Fig 3B.
• Rossner P Jr, Binkova B, Milcova A et al. Air pollution by carcinogenic PAHs and plasma levels of p53 and p21(WAF1) proteins. Mutat Res. 2007 Jul 1 [PMID: 17408702] (Human)
  
  Details:
  Direct ELISA (human blood plasma), CLONE SPECIFC Tables 1-3. Note: The specificity of the p21 antibody for p21 was validated with recombinant p21 by direct ELISA in Tables 1-3.
• Scott MT, Ingram A, Ball KL. PDK1-dependent activation of atypical PKC leads to degradation of the p21 tumour modifier protein. EMBO J. 2002 Dec 16 [PMID: 12485998] (WB)
  
  Details:
  WB: (HeLa), Fig 3D.
• Rossner P Jr, Binkova B, Chvatalova I et al. Acrylonitrile exposure: the effect on p53 and p21(WAF1) protein levels in the blood plasma of occupationally exposed workers and in vitro in human diploid lung fibroblasts. Mutat Res. 2002 May 27 [PMID: 12034325] (WB)
  
  Details:
  Direct ELISA (human blood plasma), Tables 3-5; WB (primary human lung fibroblasts), Fig 3. The specificity of the p21 antibody for p21 was validated with recombinant p21 by direct ELISA in Tables 3-5.
• Blaydes JP, Luciani MG, Pospisilova S et al. Stoichiometric phosphorylation of human p53 at Ser315 stimulates p53-dependent transcription. J Biol Chem. 2001 Feb 16 [PMID: 11078726] (WB)
  
  Details:
  WB (SAOS), Fig 5.
• Uy D, White ET, Oka T. Observation of Delta|k - l | = 3 Transitions in the nu3 Band of H3 O+ J Mol Spectrosc. 1997 Jun [PMID: 9252294] (WB)
  
  Details:
  WB (NIH3T3, MEF, ES EPL cells), Figs 1D, 3A.

FAQs for p21/CIP1/CDKN1A Antibody (NBP2-29463). (Showing 1 - 1 of 1 FAQs).

1. I would like to know the concentration of NBP2-29463.
   • NBP2-29463 (p21/CIP1/CDKN1A Antibody (WA-1 (HJ21)) has the concentration of 0.2 mg/ml.

Bioinformatics

• Gene Symbol: CDKN1A
• Uniprot:
  • P38936()