Rabbit CHK2 Polyclonal Antibody | anti-CHK2 antibody

Phospho-CHK2 (Ser33+Ser35) Antibody

Cat. Num. AAA9614109

• Gene Names: CHEK2; CDS1; CHK2; LFS2; RAD53; hCds1; HuCds1; PP1425
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Bovine (82%)
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: CHK2; Polyclonal Antibody; Phospho-CHK2 (Ser33+Ser35) Antibody; CDS 1; Cds1; Cds1 homolog; Checkpoint kinase 2; Checkpoint like protein CHK2; CHEK 2; Chek2; Chk 2; CHK2 checkpoint homolog (S. pombe); CHK2 checkpoint homolog; CHK2_HUMAN; hCds1; HuCds 1; LFS 2; LFS2; PP1425; RAD 53; RAD53; Rad53 homolog; Serine/threonine protein kinase Chk2; Serine/threonine-protein kinase Chk2; anti-CHK2 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Bovine (82%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-CHK2 (Ser33+Ser35) Antibody detects endogenous levels of CHK2 only when phosphorylated at Ser33+Ser35.; Tissue Specificity: High expression is found in testis, spleen, colon and peripheral blood leukocytes. Low expression is found in other tissues.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-CHK2 antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IHC: 1:50-1:200; IF/ICC: 1:100-1:500; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human CHK2 around the phosphorylation site of Ser33+Ser35.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: Phosphorylated. Phosphorylated at Ser-73 by PLK3 in response to DNA damage, promoting phosphorylation at Thr-68 by ATM and the G2/M transition checkpoint. Phosphorylation at Thr-68 induces homodimerization. Autophosphorylates at Thr-383 and Thr-387 in the T-loop/activation segment upon dimerization to become fully active and phosphorylate its substrates like for instance CDC25C. DNA damage-induced autophosphorylation at Ser-379 induces CUL1-mediated ubiquitination and regulates the pro-apoptotic function. Phosphorylation at Ser-456 also regulates ubiquitination. Phosphorylated by PLK4.Ubiquitinated. CUL1-mediated ubiquitination regulates the pro-apoptotic function. Ubiquitination may also regulate protein stability. Ubiquitinated by RNF8 via 'Lys-48'-linked ubiquitination.
• Subunit Structure: Homodimer. Homodimerization is part of the activation process but the dimer may dissociate following activation. Interacts with PML. Interacts with TP53. Interacts with RB1; phosphorylates RB1. Interacts with BRCA1. Interacts (phosphorylated at Thr-68) with MDC1; requires ATM-mediated phosphorylation of CHEK2. Interacts with TP53BP1; modulates CHEK2 phosphorylation at Thr-68 in response to ionizing radiation. Interacts with CDC25A; phosphorylates CDC25A and mediates its degradation in response to ionizing radiation. Interacts with CUL1; mediates CHEK2 ubiquitination and regulation. Interacts with CDKN2AIP. Interacts (via protein kinase domain) with CCAR2 (via N-terminus). Interacts with SIRT1.
• Similarity: Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. CHK2 subfamily.
• Subcellular Location: Nucleus.; Note: Isoform 10 is present throughout the cell.Nucleus. Nucleus. Nucleus. Nucleus. Nucleus>PML body. Nucleus>Nucleoplasm.; Note: Recruited into PML bodies together with TP53.
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis of extracts from Mouse heart, using Phospho-CHK2 (Ser33+Ser35) Antibody. The lane on the left was treated with blocking peptide.)

Western Blot (WB)

(Western blot analysis of CHK2 (Phospho-Ser33+Ser35) using Jurkat whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunohistochemistry (IHC)

(At 1/200 staining Rat lung tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/200 staining Mouse heart tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/200 staining Human kidney tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The tissue was then blocked and incubated with the antibody for 1.5 hours at 22 degree C. An HRP conjugated goat anti-rabbit antibody was used as the secondary antibody.)

Testing Data

(At 25 degree C. The primary antibody was diluted at 1/200 and incubated with the sample for 1 hour at 37 degree C. An Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) Ab, diluted at 1/600, was used as the secondary antibody.)

Related Product Information for anti-CHK2 antibody

Serine/threonine-protein kinase which is required for checkpoint-mediated cell cycle arrest, activation of DNA repair and apoptosis in response to the presence of DNA double-strand breaks. May also negatively regulate cell cycle progression during unperturbed cell cycles. Following activation, phosphorylates numerous effectors preferentially at the consensus sequence [L-X-R-X-X-S/T]. Regulates cell cycle checkpoint arrest through phosphorylation of CDC25A, CDC25B and CDC25C, inhibiting their activity. Inhibition of CDC25 phosphatase activity leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. May also phosphorylate NEK6 which is involved in G2/M cell cycle arrest. Regulates DNA repair through phosphorylation of BRCA2, enhancing the association of RAD51 with chromatin which promotes DNA repair by homologous recombination. Also stimulates the transcription of genes involved in DNA repair (including BRCA2) through the phosphorylation and activation of the transcription factor FOXM1. Regulates apoptosis through the phosphorylation of p53/TP53, MDM4 and PML. Phosphorylation of p53/TP53 at 'Ser-20' by CHEK2 may alleviate inhibition by MDM2, leading to accumulation of active p53/TP53. Phosphorylation of MDM4 may also reduce degradation of p53/TP53. Also controls the transcription of pro-apoptotic genes through phosphorylation of the transcription factor E2F1. Tumor suppressor, it may also have a DNA damage-independent function in mitotic spindle assembly by phosphorylating BRCA1. Its absence may be a cause of the chromosomal instability observed in some cancer cells. Promotes the CCAR2-SIRT1 association and is required for CCAR2-mediated SIRT1 inhibition.

NCBI and Uniprot Product Information

• NCBI GI #: 54112407
• NCBI GeneID: 11200
• NCBI Accession #: NP_001005735.1
• NCBI GenBank Nucleotide #: NM_001005735.1
• UniProt Accession #: O96017; Q6QA03; Q6QA04; Q6QA05; Q6QA06; Q6QA07; Q6QA08; Q6QA10; A8K3Y9; B7ZBF3; B7ZBF4; B7ZBF5
• Molecular Weight: 60,915 Da
• NCBI Official Full Name: serine/threonine-protein kinase Chk2 isoform c
• NCBI Official Synonym Full Names: checkpoint kinase 2
• NCBI Official Symbol: CHEK2
• NCBI Official Synonym Symbols: CDS1; CHK2; LFS2; RAD53; hCds1; HuCds1; PP1425
• NCBI Protein Information: serine/threonine-protein kinase Chk2; cds1 homolog; CHK2 checkpoint homolog; checkpoint-like protein CHK2
• UniProt Protein Name: Serine/threonine-protein kinase Chk2
• UniProt Gene Name: CHEK2
• UniProt Synonym Gene Names: CDS1; CHK2; RAD53; Hucds1; hCds1
• UniProt Entry Name: CHK2_HUMAN

NCBI Description

In response to DNA damage and replication blocks, cell cycle progression is halted through the control of critical cell cycle regulators. The protein encoded by this gene is a cell cycle checkpoint regulator and putative tumor suppressor. It contains a forkhead-associated protein interaction domain essential for activation in response to DNA damage and is rapidly phosphorylated in response to replication blocks and DNA damage. When activated, the encoded protein is known to inhibit CDC25C phosphatase, preventing entry into mitosis, and has been shown to stabilize the tumor suppressor protein p53, leading to cell cycle arrest in G1. In addition, this protein interacts with and phosphorylates BRCA1, allowing BRCA1 to restore survival after DNA damage. Mutations in this gene have been linked with Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype usually associated with inherited mutations in TP53. Also, mutations in this gene are thought to confer a predisposition to sarcomas, breast cancer, and brain tumors. This nuclear protein is a member of the CDS1 subfamily of serine/threonine protein kinases. Several transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Apr 2012]

Uniprot Description

Chk2: a CAMK protein kinase of the Rad53 family. A cell cycle checkpoint regulator and putative tumor suppressor. It contains a forkhead-associated protein interaction domain essential for activation in response to DNA damage and is rapidly phosphorylated in response to replication blocks and DNA damage. Phosphorylates and inhibits CDC25C, preventing entry into mitosis, p53, leading to cell cycle arrest in G1, and BRCA1, restoring survival after DNA damage. Twelve splice variant isoforms have been described for human Chk2. LOF mutants cause Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype also caused by p53 mutations. Familial mutations also associated with prostate and breast cancer, and mutations also seen in a variety of sporadic cancers and cell lines.

Protein type: Protein kinase, CAMK; Tumor suppressor; EC 2.7.11.1; Protein kinase, Ser/Thr (non-receptor); Kinase, protein; CAMK group; RAD53 family

Chromosomal Location of Human Ortholog: 22q12.1

Cellular Component: nucleoplasm; Golgi apparatus; chromosome, telomeric region; PML body

Molecular Function: identical protein binding; protein serine/threonine kinase activity; protein binding; protein homodimerization activity; metal ion binding; ubiquitin protein ligase binding; protein kinase binding; ATP binding

Biological Process: DNA damage induced protein phosphorylation; protein stabilization; transcription, DNA-dependent; protein amino acid autophosphorylation; positive regulation of transcription, DNA-dependent; DNA damage response, signal transduction; protein amino acid phosphorylation; DNA damage response, signal transduction resulting in induction of apoptosis; regulation of transcription, DNA-dependent; cell division; cellular protein catabolic process; double-strand break repair; DNA damage checkpoint; response to gamma radiation; regulation of protein catabolic process; G2/M transition of mitotic cell cycle; response to DNA damage stimulus

Disease: Li-fraumeni Syndrome 2; Prostate Cancer; Breast Cancer; Osteogenic Sarcoma