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CHK2 phospho T68 Antibody

Rabbit Polyclonal
NCI Collaboration
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Western blot using Rockland's Affinity Purified anti-Chk2 pT68 antibody shows detection of a predominant band at ~60 kDa corresponding to phosphorylated Chk2 (arrowhead) in MCF-7 whole cell lysates after treatment with doxorubicin.  Chk2 phosphorylation was induced using increasing concentrations of the DNA damaging agent doxorubicin as indicated for 24 h prior to lysate production. Personal communication, Xiao HeYang, University of Oklahoma Health Sciences Center.
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Number:
600-401-280
Size:
100 µg
Price:
$309.00
Shipping:
$20.00
Handling:
$20.00
Availability:
Ships next business day.

CHK2 phospho T68 Antibody Properties

Description
Anti-CHK2 pT68 (RABBIT) Antibody - 600-401-280
Host
Rabbit
Target Species
Human
Known Cross Reactivity
human
Clonality
Polyclonal
Application
ELISA : 1:10,000 - 1:70,000
Western Blot : 1:200 - 1:2,000
ImmunoPrecipitation: 1:100
Other Dilution: User Optimized
Physical State
Liquid (sterile filtered)
Shipping Condition
Dry Ice
Format
IgG
Label
Concentration
1.0mg/ml by UV absorbance at 280 nm
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% (w/v) Sodium Azide
Stabilizer
None
GeneName
CHEK2
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CHK2 phospho T68 Antibody Description

Background
CHK2 (also known as CHEK2, Protein kinase CHK2 isoform a, and checkpoint-like protein) is a serine/ threonine-protein kinase involved in the control of cell cycle checkpoints and may also participate in transduction of the DNA damage and replicational stress signals. CHK2 is the mammalian ortholog of the budding yeast Rad53 and fission yeast Cds1 checkpoint kinases. The amino-terminal domain of CHK2 contains a series of seven serine and threonine residues (Ser19, Thr26, Ser28, Ser33, Ser35, Ser50 and Thr68) followed by glutamine (SQ or TQ motif). These are known to be preferred sites for phosphorylation by ATM/ATR kinases. Indeed, after DNA damage by ionizing radiation (IR), UV irradiation or hydroxyurea treatment, Thr68 and other sites in this region become phosphorylated by ATM/ATR.  The SQ/TQ cluster domain, therefore, seems to have a regulatory function.  Phosphorylation at Thr68 is a prerequisite for the subsequent activation step, which is attributable to autophosphorylation of Chk2 on residues Thr383 and Thr387 in the activation loop of the kinase domain.  CHK2 inhibits CDC25C phosphatase by phosphorylating it on Ser-216, preventing the entry into mitosis. This kinase may have a role in meiosis as well. Kinase activity is up regulated by autophosphorylation and the protein is rapidly phosphorylated in response to DNA damage and to replication block.
Synonyms
Serine/threonine-protein kinase Chk2 EC=2.7.11.1 Cds1
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding aa 64-73 of Human CHK2.
Immunogen Type
Peptide
Post Translational Modification
Phosphorylation
Storage Condition
Store vial at -20° C prior to opening.  Aliquot contents and freeze at -20° C or below for extended storage.  Avoid cycles of freezing and thawing.  Centrifuge product if not completely clear after standing at room temperature.  This product is stable for several weeks at 4° C as an undiluted liquid.  Dilute only prior to immediate use.
Application Note
This affinity purified antibody has been tested for use in ELISA and by western blot.  Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 61 kDa in size corresponding to CHK2 by western blotting in the appropriate cell lysate or extract.  Less than 1% reactivity is observed against the non-phosphorylated form of the immunizing peptide.  This antibody is phospho specific for pT68 of CHK2.
Purity/Specifity
This affinity purified antibody is directed against the phosphorylated form of human CHK2 at the pT68 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification.  Antiserum was first purified against the phosphorylated form of the immunizing peptide.  The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.  Reactivity occurs against human CHK2 pT68 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human CHK2 is minimal by ELISA. The antibody does not cross-react with Chk2 phosphorylated at other sites.  A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence.  Cross reactivity with CHK2 protein from mouse and rat may occur as sequence homology varies by one amino acid residues in this sequence (90% homology). Cross reactivity with CHK2 homologues from other sources has not been determined.
Disclaimer Note-General
This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information. All products of animal origin manufactured by Rockland Immunochemicals are derived from starting materials of North American origin. Collection was performed in United States Department of Agriculture (USDA) inspected facilities and all materials have been inspected and certified to be free of disease and suitable for exportation. All properties listed are typical characteristics and are not specifications. All suggestions and data are offered in good faith but without guarantee as conditions and methods of use of our products are beyond our control. All claims must be made within 30 days following the date of delivery. The prospective user must determine the suitability of our materials before adopting them on a commercial scale. Suggested uses of our products are not recommendations to use our products in violation of any patent or as a license under any patent of Rockland Immunochemicals, Inc. If you require a commercial license to use this material and do not have one, then return this material, unopened to: Rockland Inc., P.O. BOX 5199, Limerick, Pennsylvania, USA.
General Reference
Blasina,A., de Weyer,I.V., Laus,M.C., Luyten,W.H., Parker,A.E. and McGowan,C.H. (1999) A human homologue of the checkpoint kinase Cds1 directly inhibits Cdc25 phosphatase. Curr. Biol. 9 (1), 1-10. Matsuoka,S., Huang,M. and Elledge,S.J. (1998) Linkage of ATM to cell cycle regulation by the Chk2 protein kinase. Science 282 (5395), 1893-1897. Brown,A.L., Lee,C.H., Schwarz,J.K., Mitiku,N., Piwnica-Worms,H. and Chung,J.H. (1999) A human Cds1-related kinase that functions downstream of ATM protein in the cellular response to DNA damage. Proc. Natl. Acad. Sci. U.S.A. 96 (7), 3745-3750.
Specific Reference
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Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC
Reacts With
human
Catalog Number
100-401-A17

Product Type
Primary Antibodies;
Applications
ELISA, WB, IHC, IP, ChIP
Reacts With
human
Catalog Number
200-301-174

Product Type
Secondary Antibodies;
Applications
FLISA, IF, FC
Catalog Number
211-1202

Product Type
Primary Antibodies;
Applications
ELISA, WB
Reacts With
human, mouse, bovine, dog, macaque, chimpanzee, rat
Catalog Number
600-401-859

Product Label

 

Description
Unconjugated
Color
Conjugation Reference
Molecular Weight
Excitation Wavelength
Conjugation Chemistry