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Details for Product No. ABIN487338

Cyclin-Dependent Kinase Inhibitor 2D (p19, Inhibits CDK4) (CDKN2D) antibody

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Antigen
Synonyms p19, INK4D, p19-INK4D, INK4d, p19INK4d, ink4d, p19-ink4d, CDKN2D, ank2
Reactivity
»Alternatives Human
Host
»Alternatives Mouse
Clonality (Clone) Monoclonal ()
Conjugate
»Alternatives Un-conjugated
Application
»Alternatives Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
Pubmed 1 reference available
Catalog no. ABIN487338
Quantity 0.1 mg
Price
478.50 $   Plus shipping costs $45.00
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Availability Will be delivered in 6 to 8 Business Days
Immunogen Bacterially produced GST-tagged fusion proteins of full-length p19INK4d. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.
Clone DCS-100
Isotype IgG1
Characteristics Synonyms: p19-INK4d, p19, INK4D, Cyclin-dependent kinase 4 inhibitor D
Purification Protein-A Sepharose Chromatography.
Alternative Name CDKN2D / p19INK4d
Background The INK4 family of proteins consists of four members that block progression from theG(1)-to-S phase of the cell cycle by inhibiting the activity of Cdk4 and Cdk6. p19INK4d is a165 amino acid protein with strong structural and functional similarity to p16INK4a, aknown tumor suppressor. Mutations in p19INK4d have also been associated with humanosteosarcomas.
Gene ID 1032
UniProt P55273
Application Notes Western Blot: 1 µg/mlPositive Control: Jurkat cells. Immunoprecipitation: 3 µg/200-300 µl of cell extract. Positive Control: Jurkat 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 Control: 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-p19INK4d (DCS-100) monoclonal antibody (1μg/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 HRP-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: Jurkat Cells. 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-p19INK4d (DCS-100) 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: Jurkat cells.
Restrictions For Research Use only
Format Liquid
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
Background publications Thullberg, Welcker, Bartkova et al.: "Monoclonal antibody probes for p21WAF1/CIP1 and the INK4 family of cyclin-dependent kinase inhibitors." in: Hybridoma, Vol. 19, Issue 1, pp. 63-72, 2000 (PubMed).

Alternatives for antigen "Cyclin-Dependent Kinase Inhibitor 2D (p19, Inhibits CDK4) (CDKN2D)", type "Antibodies"
Hosts (25), (20)
Reactivities (46), (13), (12)
Applications (25), (14), (13), (13), (10), (7), (4), (4), (3), (3), (2), (1), (1)
Conjugates (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Epitopes (5), (1), (1), (1)
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