Product Pathways - Cell Cycle / Checkpoint
UV Induced DNA Damage Response Antibody Sampler Kit #8342
|8342S||1 Kit (7 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|Phospho-Histone H2A.X (Ser139) (20E3) Rabbit mAb #9718||40 µl||W, IHC-P, IF-IC, F||H, M, R, Mk||15||Rabbit IgG|
|Phospho-cdc25C (Ser216) (63F9) Rabbit mAb #4901||40 µl||W, IP, IHC-P||H, Mk||60||Rabbit IgG|
|Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348||40 µl||W, IF-IC, F||H, M, R, Mk||56||Rabbit IgG|
|RPA32/RPA2 (4E4) Rat mAb #2208||40 µl||W, IP, IF-IC||H, M, R, Hm, Mk||32||Rat IgG1|
|ATRIP Antibody #2737||40 µl||W, IP, IF-IC||H||82 (isoform 1)||Rabbit|
|Phospho-ATR (Ser428) Antibody #2853||40 µl||W||H, M, R, Mk||300||Rabbit|
|Microcephalin-1/BRIT1 (D38G5) Rabbit mAb #4120||40 µl||W||H, M, R, Mk||100||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-rat IgG, HRP-linked Antibody #7077||100 µl||Goat|
Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, IP=Immunoprecipitation
Reactivity Key: H=Human, M=Mouse, R=Rat, Mk=Monkey, Hm=Hamster
Western blot analysis of extracts from various cell lines using RPA32 (4E4) Rat mAb #2208.
Western blot analysis of extracts from various cell lines, untreated or UV-treated, using Phospho-Chk1 (Ser345) (133D3) Rabbit mAb #2348.
Western blot analysis of extracts from Jurkat and HEL cells using ATRIP Antibody #2737.
Western blot analysis of Raw 264.7, SV-T2, and HT-29 cells, untreated or UV-treated (50 mJ, 30 min), using Phospho-ATR (Ser428) Antibody #2853. λ phosphatase (10,000 Units/ml, 1hr) was used to demonstrate the phospho-specificity of the antibody.
Western blot analysis of extracts from various cell lines using Microcephalin-1/BRIT1 (D38G5) Rabbit mAb #4120.
Western blot analysis of HT-29 cells, untreated, nocodazole-treated, and λ phosphotase-treated, using Phospho-cdc25C (Ser216) (63F9) Rabbit mAb #4901 (upper) or cdc25C (5H9) Rabbit mAb #4688 (lower).
The UV Induced DNA Damage Response Antibody Sampler Kit offers an economical means of investigating proteins involved in the cellular response to UV-induced DNA damage. The kit contains enough primary and secondary antibody to perform four western blot experiments per primary.
Specificity / Sensitivity
Antibodies detect endogenous levels of the respective target proteins.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly267 of human ATRIP protein or Ser428 of human ATR protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with a recombinant full-length human Maltose Binding Protein-RPA32 fusion protein or a synthetic peptide corresponding to residues surrounding Tyr415 of human Microcephalin-1/BRIT1 protein. Activation state monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser139 of human H2A.X protein, Ser216 of human cdc25C protein, or Ser345 of human Chk1 protein, respectively.
Exposure to ultraviolet radiation (UV) has a profound impact on human health and disease (1). Low level UV exposure induces the production of vitamin D and is a key regulator of calcium metabolism. Conversely, overexposure to UV is associated with an increased risk of cancer, immunosuppression, and many eye disorders, such as cataracts. Photons of UV light can directly damage DNA causing thymine dimers and other pyrimidine dimers between adjacent bases (2). Free radicals and reactive oxygen species induced by UV exposure also result in DNA lesions and have been linked to malignant melanoma (3). DNA damage from replicative stress and genotoxic agents like UV activate the ATR-mediated checkpoint pathway and stimulate DNA repair, cell cycle arrest, and apoptosis (4). ATR recruitment to sites of DNA damage and activation depends, at least in part, on interaction with the complex of single-stranded DNA, Replication Protein A (RPA), and direct binding to the ATR-associated adapter protein, ATRIP (5). In addition, the Rad17-RFC and Rad9-Rad1-Hus1 (9-1-1) protein complexes are independently recruited with TopBP1 to fully activate the checkpoint response (6,7). BRIT1 (MCPH1) is required for UV-induced formation of ATR, RPA, and p-Rad17 foci at sites of DNA damage (8-10) and may regulate the expression of several DNA damage response proteins (11). Once activated, ATR phosphorylates a number of mediators, including histone H2AX Ser139 and Chk1 kinase at Ser345. H2AX phosphorylation is a marker of DNA damage. Complete loss of H2AX results in reduced Chk1 activation and impaired survival of cells after UV exposure (12). Chk1 and Chk2 kinase activation is essential for checkpoint-mediated control of cell cycle progression (4). Checkpoint kinases stimulate cell cycle arrest by phosphorylation of a group of tyrosine phosphatases known as Cdc25A, Cdc25B, and Cdc25C (13 -15). Both Chk1 and Chk2 kinases phosphorylate Cdc25C at Ser216 in response to DNA damage and stimulate arrest (16-17).
- von Thaler, A.K. et al. (2010) Exp Dermatol 19, 81-8.
- Rastogi, R.P. et al. (2010) J Nucleic Acids 2010, 592980.
- Narayanan, D.L. et al. (2010) Int J Dermatol 49, 978-86.
- Zhou, B.B. and Elledge, S.J. (2000) Nature 408, 433-9.
- Zou, L. and Elledge, S.J. (2003) Science 300, 1542-8.
- Zou, L. et al. (2002) Genes Dev 16, 198-208.
- Mordes, D.A. and Cortez, D. (2008) Cell Cycle 7, 2809-12.
- Rai, R. et al. (2006) Cancer Cell 10, 145-57.
- Peng, G. et al. (2009) Nat Cell Biol 11, 865-72.
- Lin, S.Y. et al. (2010) Yonsei Med J 51, 295-301.
- Lin, S.Y. et al. (2005) Proc Natl Acad Sci U S A 102, 15105-9.
- Revet, I. et al. (2011) Proc Natl Acad Sci U S A 108, 8663-7.
- Mailand, N. et al. (2000) Science 288, 1425-9.
- Sanchez, Y. et al. (1997) Science 277, 1497-501.
- Matsuoka, S. et al. (1998) Science 282, 1893-7.
- Blasina, A. et al. (1999) Curr Biol 9, 1-10.
- Furnari, B. et al. (1999) Mol Biol Cell 10, 833-45.
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For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.
Select rabbit monoclonal antibodies are developed, validated, and produced at CST using in part technology under license (granting certain rights including those under U.S. Patents No. 5,675,063 and in some instances 7,429,487) from Epitomics, Inc.