p21 Antikörper

Produktnummer ABIN487485

Produktdetails anti-p21 Antikörper

Target: p21 (CDKN1A) (Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A))

Reaktivität: Human

Wirt: Maus

Klonalität: Monoklonal

Konjugat: Dieser p21 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)

Spezifität: This antibody reacts with Human p21WAF1/CIP1.

Kreuzreaktivität (Details): Species reactivity (tested):Human.

Produktmerkmale: Synonyms: CAP20, CDKN1, CIP1, MDA6, MDA-6, PIC1, SDI1, WAF1, Cyclin-dependent kinase inhibitor 1,CDK-interacting protein 1, Melanoma differentiation-associated protein 6

Aufreinigung: 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

Klon: DCS-60

Isotyp: IgG2a

Anwendungsinformationen

Applikationshinweise: 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.

Protokoll: 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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Konzentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Konservierungsmittel: Without preservative

Lagerung: -20 °C

Informationen zur Lagerung: Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch.

Haltbarkeit: 12 months

Details zu p21

Target: p21 (CDKN1A) (Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A))

Andere Bezeichnung: CDKN1A / p21WAF1 (CDKN1A Produkte)

Synonyme: CCND1 antikoerper, CDKN1A antikoerper, CAP20 antikoerper, CDKN1 antikoerper, CIP1 antikoerper, MDA-6 antikoerper, P21 antikoerper, SDI1 antikoerper, WAF1 antikoerper, p21CIP1 antikoerper, CDKI antikoerper, Cdkn1 antikoerper, Waf1 antikoerper, mda6 antikoerper, p21Cip1 antikoerper, p21WAF antikoerper, Cip1 antikoerper, UV96 antikoerper, p21 antikoerper, cip1 antikoerper, p16Xic2 antikoerper, sdi1 antikoerper, waf1 antikoerper, cap20 antikoerper, cdkn1 antikoerper, mda-6 antikoerper, p21cip1 antikoerper, si:ch1073-48m5.2 antikoerper, KRAS2 antikoerper, p21ras antikoerper, cyclin D1 antikoerper, cyclin dependent kinase inhibitor 1A antikoerper, cyclin-dependent kinase inhibitor 1A (P21) antikoerper, cyclin-dependent kinase inhibitor 1A antikoerper, cyclin-dependent kinase inhibitor 1A L homeolog antikoerper, cyclin-dependent kinase inhibitor 1 antikoerper, KRAS proto-oncogene, GTPase antikoerper, CCND1 antikoerper, CDKN1A antikoerper, Cdkn1a antikoerper, cdkn1a.L antikoerper, cki1 antikoerper, cdkn1a antikoerper, KRAS antikoerper

Hintergrund: 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

Gen-ID: 1026

UniProt: P38936

Pathways: p53 Signalweg, PI3K-Akt Signalweg, Zellzyklus, AMPK Signaling, Fc-epsilon Rezeptor Signalübertragung, EGFR Signaling Pathway, Neurotrophin Signalübertragung, Mitotic G1-G1/S Phases, DNA Replication, Hepatitis C, Synthesis of DNA, Autophagie