Phone:
+1 877 302 8632
Fax:
+1 888 205 9894 (Toll-free)
E-Mail:
orders@antibodies-online.com

p21 antibody

CDKN1A Reactivity: Human WB, IHC (p), IP Host: Mouse Monoclonal DCS-60 unconjugated
Catalog No. ABIN487485
  • Target See all p21 (CDKN1A) Antibodies
    p21 (CDKN1A) (Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A))
    Reactivity
    • 250
    • 117
    • 94
    • 36
    • 14
    • 12
    • 5
    • 4
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Host
    • 216
    • 54
    • 1
    Mouse
    Clonality
    • 209
    • 61
    • 1
    Monoclonal
    Conjugate
    • 112
    • 20
    • 18
    • 18
    • 13
    • 11
    • 6
    • 6
    • 6
    • 6
    • 6
    • 6
    • 5
    • 5
    • 5
    • 5
    • 5
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    This p21 antibody is un-conjugated
    Application
    • 180
    • 109
    • 67
    • 66
    • 66
    • 66
    • 62
    • 43
    • 41
    • 33
    • 19
    • 16
    • 15
    • 5
    • 2
    • 2
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
    Specificity
    This antibody reacts with Human p21WAF1/CIP1.
    Cross-Reactivity (Details)
    Species reactivity (tested):Human.
    Characteristics
    Synonyms: CAP20, CDKN1, CIP1, MDA6, MDA-6, PIC1, SDI1, WAF1, Cyclin-dependent kinase inhibitor 1,CDK-interacting protein 1, Melanoma differentiation-associated protein 6
    Purification
    Protein-A Sepharose Chromatography.
    Immunogen
    Full-length Recombinant Human p21WAF1/CIP1. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte
    Clone
    DCS-60
    Isotype
    IgG2a
    Top Product
    Discover our top product CDKN1A Primary Antibody
  • Application Notes
    Western Blot: 1-5 μg/mLPositive Control: HeLa Cells. Immunoprecipitation: 3 μg/200-300 μL of cell extract. Positive Control: HeLa Cells. Immunohistochemistry: 1-5 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Microwave oven: 2 times for 10 minutes each in citrate buffer ( pH 6.5). Positive Controls: Tonsil Tissue. Detailed procedure is provided in Protocols.
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.
    Protocol
    SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for the transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with the anti-p21WAF1/CIP1 (DCS-60) monoclonal antibody (1-5μg/mL) diluted with 1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriatechemiluminescence reagents for 1 minute. Remove extra reagent from the membrane bydabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Theconditions for exposure and development may vary. Positive Control for Western blotting: HeLa. Immunoprecipitation1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. 3) Add 3 μg of the anti-p21WAF1/CIP1 (DCS-60) monoclonal antibody into 250 μL of thesupernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4°C. Add 20μL of 50% Protein A-agarose beads resuspended in the Lysis buffer. Mix well and incubatewith gentle agitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for10 seconds). 5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting. )Positive Controls for immunoprecipitation: HeLa cells.
    Restrictions
    For Research Use only
  • Concentration
    1.0 mg/mL
    Buffer
    PBS, pH 7.2 containing 50 % Glycerol without preservatives.
    Preservative
    Without preservative
    Storage
    -20 °C
    Storage Comment
    Store the antibody undiluted at -20 °C.
    Shelf life: one year from despatch.
    Expiry Date
    12 months
  • Target
    p21 (CDKN1A) (Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A))
    Alternative Name
    CDKN1A / p21WAF1 (CDKN1A Products)
    Synonyms
    CCND1 antibody, CDKN1A antibody, CAP20 antibody, CDKN1 antibody, CIP1 antibody, MDA-6 antibody, P21 antibody, SDI1 antibody, WAF1 antibody, p21CIP1 antibody, CDKI antibody, Cdkn1 antibody, Waf1 antibody, mda6 antibody, p21Cip1 antibody, p21WAF antibody, Cip1 antibody, UV96 antibody, p21 antibody, cip1 antibody, p16Xic2 antibody, sdi1 antibody, waf1 antibody, cap20 antibody, cdkn1 antibody, mda-6 antibody, p21cip1 antibody, si:ch1073-48m5.2 antibody, KRAS2 antibody, p21ras antibody, cyclin D1 antibody, cyclin dependent kinase inhibitor 1A antibody, cyclin-dependent kinase inhibitor 1A (P21) antibody, cyclin-dependent kinase inhibitor 1A antibody, cyclin-dependent kinase inhibitor 1A L homeolog antibody, cyclin-dependent kinase inhibitor 1 antibody, KRAS proto-oncogene, GTPase antibody, CCND1 antibody, CDKN1A antibody, Cdkn1a antibody, cdkn1a.L antibody, cki1 antibody, cdkn1a antibody, KRAS antibody
    Background
    P21WAF1/CIP1 is one member of the CIP/KIP family that controls the cell cycle by inhibiting cyclin dependent kinases (CDKs) activity. Increased expression of p21WAF1/CIP1 may play an important role in the growth arrest induced in transformed cells. It is reported that hypermethylation of the p21WAF1/CIP1 promoter region inactivate p21WAF1/CIP1 gene leading tumorigenesis, and also reported that p21WAF1/CIP1 acts as an inhibitor of apoptosis in a number of systems in addition to being an inhibitor of cell proliferation.Synonyms: CAP20, CDK-interacting protein 1, CDKN1, CIP1, Cyclin-dependent kinase inhibitor 1, MDA-6, MDA6, Melanoma differentiation-associated protein 6, PIC1, SDI1, WAF1
    Gene ID
    1026
    UniProt
    P38936
    Pathways
    p53 Signaling, PI3K-Akt Signaling, Cell Division Cycle, AMPK Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Mitotic G1-G1/S Phases, DNA Replication, Hepatitis C, Synthesis of DNA, Autophagy
You are here:
Support