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Purified Mouse Anti-p21
Purified Mouse Anti-p21
Western blot analysis of p21 in human and mouse. MCF-7 human (lanes 1 and 2) and EL4 mouse (lanes 3 and 4) lysates were incubated with clone SXM30 (lanes 1 and 3) or a mouse IgG1 isotype (negative) control (lanes 2 and 4). The mouse (Ms) homolog of human (Hu) p21 is slightly smaller than that human p21.
Purified Mouse Anti-p21

Anti-human p21, Cat. No. 556431. Formalin-fixed, paraffin-embedded tissue section of human colon was stained with anti-p21 (clone SXM30), followed by a DAB chromogen detection system, then counter stained with Hematoxylin. Antigen retrieval using BD Pharmingen™ Retrievagen A Buffer Set (Cat. No. 550524) is required. Positive staining can be identified as the intensed brown labeling.

Western blot analysis of p21 in human and mouse. MCF-7 human (lanes 1 and 2) and EL4 mouse (lanes 3 and 4) lysates were incubated with clone SXM30 (lanes 1 and 3) or a mouse IgG1 isotype (negative) control (lanes 2 and 4). The mouse (Ms) homolog of human (Hu) p21 is slightly smaller than that human p21.

Anti-human p21, Cat. No. 556431. Formalin-fixed, paraffin-embedded tissue section of human colon was stained with anti-p21 (clone SXM30), followed by a DAB chromogen detection system, then counter stained with Hematoxylin. Antigen retrieval using BD Pharmingen™ Retrievagen A Buffer Set (Cat. No. 550524) is required. Positive staining can be identified as the intensed brown labeling.

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BD Pharmingen™
Human (QC Testing), Mouse, Rat (Tested in Development)
Mouse IgG1, κ
Human p21
Western blot (Routinely Tested), Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required), Immunohistochemistry-frozen, Immunoprecipitation (Tested During Development)
21 kDa
0.5 mg/ml
AB_396415
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


推奨アッセイ手順

Clone SXM30 has been shown to be useful for immunoprecipitation (1-2 µg/1x10^6 cells), western blot analysis (1-2 µg/ml), indirect immunofluorescence microscopy of tissuecultured cells, immunohistochemistry of frozen, acetone-fixed tissue sections (1-5 µg/ml), and paraffin-embedded tissue sections requiring antigen retrieval (20 µg/ml). These applications are similar to that described for a related p21 clone, SX118.3.  MCF-7 human breast carcinoma cells (ATCC HTB-22), HL-60 human leukemia cells (ATCC CCL-240), and EL4 mouse lymphoma cells (ATCC TIB-39) are suggested as positive controls. In immunohistochemistry, SX118 staining is primarily nuclear. Tonsil and colon are suggested as positive controls for tissue staining.  In order to obtainn optimal indirect immunohistochemical staining, clone SXM30 should be titrated at 10-20 ug/ml and visualized via a three-step staining procedure in combination with a biotinylated polyclonal anti-mouse Ig (Cat. No. 550337, multiple absorbson) as the secondary antibody and Streptavidin-HRP (Cat. No. 550946) together with the DAB chromogen detection system (Cat. No. 550880).

Refer to Fredersdorf et al., for information regarding induction of p21 expression, and for p21 distribution in different types of human tissues.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
556431 Rev. 8
抗体の詳細
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SXM30

Cyclins and cyclin-dependent kinases (cdks) are evolutionarily conserved proteins that are essential for cell-cycle control in eukaryotes. Cyclins (regulatory subunits) bind to cdks (catalytic subunits) to form complexes that regulate the progression of the cell cycle. The activity of these complexes is modulated by activating and inhibitory phosphorylation events, as well as by interactions with small regulatory proteins including, p16, p21, p27, and others. These proteins, referred to as inhibitors of Cdk activity (CdkIs), bind to cyclins, cdks or their complexes. p21, also known as senescent cell-derived inhibitor 1 (Sdi1), wild-type p53-activated fragment 1 (Waf1), Cdk-interacting protein 1 (Cip1), and p53-regulated inhibitor of Cdks (Pic1) inhibits cyclin D-cdk4, cyclin E-cdk3, cyclin A-cdk2, and cyclin A-cdk1. p21 can also inhibit cell cycle progression by binding to PCNA and blocking DNA replication. p21 has also been shown to be a component of active cyclin-cdk complexes, suggesting that p21-containing complexes may shift between active and inactive states through changes in p21 content. Active, p21-containing complexes appear to contain one p21 molecule, whereas inactive compexes contain multiple p21 molecules. The expression of p21 can be induced in response to number of signals, including transcriptional upregulation by the tumor suppressor protein, p53. Human p21 has a calculated molecular weight of 18 kDa and runs at 21 kDa in SDS-PAGE.

Clone SXM30 reacts with human, rat, and mouse p21. With regard to cross-species recognition, SXM30 has similar properties to a related p21 clone, SX118. Purified, recombinant human p21-fusion protein was used as immunogen.

556431 Rev. 8
フォーマットの詳細
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
556431 Rev.8
引用&参考文献
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Development References (3)

  1. Fredersdorf S, Milne AW, Hall PA, Lu X. Characterization of a panel of novel anti-p21Waf1/Cip1 monoclonal antibodies and immunochemical analysis of p21Waf1/Cip1 expression in normal human tissues. Am J Pathol. 1996; 148(3):825-835. (Clone-specific: Immunofluorescence, Immunoprecipitation, Western blot). View Reference
  2. Grana X, Reddy EP. Cell cycle control in mammalian cells: role of cyclins, cyclin dependent kinases (CDKs), growth suppressor genes and cyclin-dependent kinase inhibitors (CKIs). Oncogene. 1995; 11(2):211-219. (Biology). View Reference
  3. Levine AJ. p53, the cellular gatekeeper for growth and division. Cell. 1997; 88(3):323-331. (Biology). View Reference
556431 Rev. 8

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.