Checkpoint Kinase 2 (CHEK2) antibody

Details for Product No. ABIN487312
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Antigen
Synonyms CHK-2, CDS1, CHK2, HuCds1, LFS2, PP1425, RAD53, hCds1, Cds1, HUCDS1, Rad53, Chk2, fa66f08, wu:fa66f08, zgc:55865
Reactivity
Human
(392), (167), (166), (55), (37), (36), (12), (7), (6), (1)
Host
Mouse
(318), (81), (1), (1)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(7), (7), (6), (6), (6), (6), (6), (6), (6), (6), (6), (1), (1), (1), (1), (1)
Application
Immunoprecipitation (IP), Immunohistochemistry (Frozen Paraffin-embedded Sections) (IHC (frpe)), Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
(313), (128), (122), (67), (60), (54), (34), (32), (22), (12), (4), (4), (1), (1)
Pubmed 4 references available
Quantity 0.1 mg
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Catalog No. ABIN487312
489.50 $
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Immunogen Full-length Human Chk2 fusion protein. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte
Clone DCS-270
Isotype IgG2a
Specificity This antibody reacts with Human Chk2.
Cross-Reactivity (Details) Species reactivity (tested):Human.
Characteristics Synonyms: CHEK2, CHEK-2, CHK-2, RAD53, Cds1, Serine/threonine-protein kinase Chk2, CHK2checkpoint homolog
Purification Protein-A Sepharose Chromatography.
Alternative Name CHK2
Background Checkpoint kinase 2 (Chk2), also known as Cds1, is a 61 kDa nuclear protein that functions as a checkpoint kinase to regulate the cell cycle progression following DNA damage. Chk2 inhibits CDC2 by inactivating CDC25, the phosphatase that normally activates CDC2. Other targets for Chk2 include the tumor suppressors BRCA1 and p53, which it stabilizes by phosphorylation of Ser20. Chk2 is itself phosphorylated and activated by ATM following DNA damage. Defects in Chk2 contribute to the development of human cancers, and implicate Chk2 as a candidate tumor suppressor and an attractive target for drug discovery.Synonyms: CHEK-2, CHEK2, CHK-2, CHK2 checkpoint homolog, Cds1, RAD53, Serine/threonine-protein kinase Chk2
Gene ID 11200
UniProt O96017
Application Notes Western Blot: 0.2 μg/mL. Positive Control: HeLa. Immunoprecipitation: 3 μg/200-300 μL of cell extract. Positive Control: HeLa. 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-Chk2 (DCS-270) monoclonal antibody (0. 2 μg/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 Controls for Western blotting: HeLaImmunoprecipitation1) 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-Chk2 (DCS-270) monoclonalantibody into 250 µL of the supernatant. Mix well and incubate with gentle agitation for30-120 minutes at 4°C. Add 20 µL of 50% Protein A-agarose beads resuspended in the
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 (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Expiry Date 12 months
Supplier Images
anti-Checkpoint Kinase 2 (CHEK2) antibody anti-Checkpoint Kinase 2 (CHEK2) antibody
anti-Checkpoint Kinase 2 (CHEK2) antibody (2) anti-Checkpoint Kinase 2 (CHEK2) antibody (Image 2)
anti-Checkpoint Kinase 2 (CHEK2) antibody (3) anti-Checkpoint Kinase 2 (CHEK2) antibody (Image 3)
Background publications Lukas, Bartkova, Latella et al.: "DNA damage-activated kinase Chk2 is independent of proliferation or differentiation yet correlates with tissue biology." in: Cancer research, Vol. 61, Issue 13, pp. 4990-3, 2001 (PubMed).

Latella, Lukas, Simone et al.: "Differentiation-induced radioresistance in muscle cells." in: Molecular and cellular biology, Vol. 24, Issue 14, pp. 6350-61, 2004 (PubMed).

Wei, Chou, Ou et al.: "TTK/hMps1 participates in the regulation of DNA damage checkpoint response by phosphorylating CHK2 on threonine 68." in: The Journal of biological chemistry, Vol. 280, Issue 9, pp. 7748-57, 2005 (PubMed).

Yoda, Xu, Onishi et al.: "Intrinsic kinase activity and SQ/TQ domain of Chk2 kinase as well as N-terminal domain of Wip1 phosphatase are required for regulation of Chk2 by Wip1." in: The Journal of biological chemistry, Vol. 281, Issue 34, pp. 24847-62, 2006 (PubMed).

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