hh Human p21/CIP1/CDKN1A Antibody AF1047: R&D Systems

Human p21/CIP1/CDKN1A Antibody

(1 citations)
(2 Reviews)
  
  • Species Reactivity
    Human
  • Specificity
    Detects human p21 in Western blots.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human p21
    Ser2-Pro164
    Accession # P38936
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    5 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
  • Immunoprecipitation
    1-2 µg/500 µg cell lysate
    MCF‑7 human breast cancer cell line, see our available Western blot detection antibodies
  • CyTOF-ready
    Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
  • Immunocytochemistry
    5-15 µg/mL
    See below
  • Intracellular Staining by Flow Cytometry
    0.25 µg/106 cells
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human p21/CIP1/CDKN1A by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 1 μM camptothecin (CPT) for 16 hours. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human p21/CIP1/CDKN1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1047), followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for p21/CIP1/CDKN1A at approximately 21 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunocytochemistry
p21/CIP1/CDKN1A in MCF‑7 Human Cell Line. p21/CIP1/CDKN1A was detected in immersion fixed MCF‑7 human breast cancer cell line treated with (left panel) or without (right panel) camptothecin using Goat Anti-Human p21/CIP1/CDKN1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1047) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
p21/CIP1/CDKN1A in Human Breast Cancer Tissue. p21/CIP1/CDKN1A was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using 1.7 µg/mL Goat Anti-Human p21/CIP1/CDKN1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1047) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Intracellular Staining by Flow Cytometry
Detection of p21/CIP1/CDKN1A in MCF‑7 Human Cell Line by Flow Cytometry. MCF‑7 human breast cancer cell line was unstimulated (light orange filled histogram) or treated with 1 μM camphtothecin for 16 hours, then stained with Goat Anti-Human p21/CIP1/CDKN1A Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF1047, dark orange filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with methanol.
Detection of Human p21/CIP1/CDKN1A by Simple WesternTM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 16 hours, loaded at 0.2 mg/mL. A specific band was detected for p21/CIP1/CDKN1A at approximately 30 kDa (as indicated) using 5 µg/mL of Goat Anti-Human p21/CIP1/CDKN1A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1047) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the
12-230 kDa separation system.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: p21/CIP1/CDKN1A
p21CIP1, also called CIP1 (CDK-interacting protein 1) and CDKN1A, is a 21 kDa Cyclin/Cyclin-dependent kinase (Cdk) inhibitor that blocks cell cycle progression from G1 to S phase in the cell cycle. Because p21 is a transcriptional target of the p53 tumor suppressor, p21 expression increases in cells that contain stabilized p53 due to genotoxic stress exposure.
  • Long Name:
    p21 Cyclin Dependent Kinase 4 Inhibitor 1A
  • Entrez Gene IDs:
    1026 (Human); 12575 (Mouse); 114851 (Rat)
  • Alternate Names:
    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
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Induction of alternative lengthening of telomeres-associated PML bodies by p53/p21 requires HP1 proteins.
    Authors: Jiang WQ, Zhong ZH, Nguyen A, Henson JD, Toouli CD, Braithwaite AW, Reddel RR
    J. Cell Biol., 2009;185(5):797-810.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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Average Rating: 4 (Based on 2 reviews)

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We have 2 reviews tested in 1 species: Human.
We have 2 reviews tested in 2 applications: Immunohistochemistry, Immunocytochemistry/Immunofluorescence.

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Images Ratings Applications Species Reviewed By Date Details
Immunohistochemistry Human p21/CIP1/CDKN1A Antibody AF1047
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Very Good
 IHC Human Anonymous 11/04/2016
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Immunohistochemistry Human p21/CIP1/CDKN1A Antibody AF1047
Immunohistochemistry: Human p21/CIP1/CDKN1A Antibody [AF1047]

Summary

ApplicationImmunohistochemistry
Sample TestedAdult lung
SpeciesHuman
Immunocytochemistry/Immunofluorescence Human p21/CIP1/CDKN1A Antibody AF1047
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Very Good
 ICC/IF Human Anonymous 10/18/2016
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Immunocytochemistry/Immunofluorescence Human p21/CIP1/CDKN1A Antibody AF1047
Immunocytochemistry/Immunofluorescence: Human p21/CIP1/CDKN1A Antibody [AF1047]

Summary

ApplicationImmunocytochemistry/Immunofluorescence
Sample TestedAdult lung
SpeciesHuman

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