p21 anticorps

N° du produit ABIN487485

Détail du produit anti-p21 anticorps

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

Reactivité: Humain

Hôte: Souris

Clonalité: Monoclonal

Conjugué: Cet anticorp p21 est non-conjugé

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

Specificité: This antibody reacts with Human p21WAF1/CIP1.

Réactivité croisée (Details): Species reactivity (tested):Human.

Attributs du produit: 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.

Immunogène: 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

Information d'application

Indications d'application: 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.

Protocole: 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

Stockage

Concentration: 1.0 mg/mL

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

Agent conservateur: Without preservative

Stock: -20 °C

Stockage commentaire: Store the antibody undiluted at -20 °C.
Shelf life: one year from despatch.

Date de péremption: 12 months

Détails sur p21

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

Autre désignation: CDKN1A / p21WAF1 (CDKN1A Produits)

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

Sujet: 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

ID gène: 1026

UniProt: P38936

Pathways: Signalisation p53, Signalisation PI3K-Akt, Cycle Cellulaire, 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