DNA damage activates ATM, which phosphorylates and activates CHK1 and CHK2/Cds1. The principal substrates of CHK1 and CHK2 are cdc25C (inhibited by phosphorylation), p53 (stabilized by phosphorylation), and BRCA1 (scatters through the nucleus following phosphorylation). Activation of p53 results in cell cycle arrest, or induction of apoptosis. Cdc25C inhibition prevents dephosphorylation of the inhibitory phosphotyrosine residue on cdc2/cdk1, thus preventing the G2/M transition. BRCA1 dispersion through the nucleus is associated with DNA repair.
Product Information
Format
Culture Supernatant
Control
K562 whole cell lysate.
Presentation
Unpurified rabbit monoclonal IgG in buffer containing 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA
Applications
Application
Please note that this product will not be available for sale after March 15, 2015. Please select one of the other antibodies against this target. This Anti-Chk1 (N-term) Antibody, clone E250, Rabbit is validated for use in WB, IC for the detection of Chk1 (N-term).
Key Applications
Immunocytochemistry
Western Blotting
Application Notes
Immunocytochemistry:
A previous lot of this antibody was used on Immunocytochemistry
Biological Information
Immunogen
Synthetic peptide directed against the N-terminus of human Chk1.
Epitope
N-Terminus
Clone
E250
Host
Rabbit
Specificity
This antibody recognizes Chk1 at the N-Terminus.
Isotype
IgG
Species Reactivity
Rat Human
Species Reactivity Note
Proven to react with human and mouse. Not expected to react with rat.
FUNCTION:Required for checkpoint mediated cell cycle arrest in response to DNA damage or the presence of unreplicated DNA. May also negatively regulate cell cycle progression during unperturbed cell cycles. Recognizes the substrate consensus sequence [R-X-X-S/T]. Binds to and phosphorylates CDC25A, CDC25B and CDC25C. Phosphorylation of CDC25A at 'Ser-178' and 'Thr-507' and phosphorylation of CDC25C at 'Ser-216' creates binding sites for 14-3-3 proteins which inhibit CDC25A and CDC25C. Phosphorylation of CDC25A at 'Ser-76', 'Ser-124', 'Ser-178', 'Ser-279' and 'Ser-293' promotes proteolysis of CDC25A. Inhibition of CDC25 activity leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. Binds to and phosphorylates RAD51 at 'Thr-309', which may enhance the association of RAD51 with chromatin and promote DNA repair by homologous recombination. Binds to and phosphorylates TLK1 at 'Ser-743', which prevents the TLK1-dependent phosphorylation of the chromatin assembly factor ASF1A. This may affect chromatin assembly during S phase or DNA repair. May also phosphorylate multiple sites within the C-terminus of TP53, which promotes activation of TP53 by acetylation and enhances suppression of cellular proliferation. Ref.1 Ref.6 Ref.9 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.20 Ref.22 Ref.24 Ref.25 Ref.27 Ref.29 Ref.33 Ref.35 Ref.36
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
SUBUNIT: Interacts with BRCA1, CLSPN, PPM1D, RAD51, TIMELESS, XPO1/CRM1 and YWHAZ/14-3-3 zeta. Ref.1 Ref.35 Ref.14 Ref.21 Ref.23 Ref.30 Ref.32 Ref.34
SUBCELLULAR LOCATION:Nucleus. Cytoplasm. Note= Nuclear export is mediated at least in part by XPO1/CRM1. Also localizes to the centrosome specifically during interphase, where it may protect centrosomal CDC2 kinase from inappropriate activation by cytoplasmic CDC25B. Ref.1 Ref.29 Ref.14 Ref.21 Ref.31
SPECIFICITY:Expressed ubiquitously with the most abundant expression in thymus, testis, small intestine and colon. Ref.1
DOMAIN: The autoinhibitory region (AIR) inhibits the activity of the kinase domain. Ref.28
PTM:Phosphorylated by ATR in a RAD17-dependent manner in response to ultraviolet irradiation and inhibition of DNA replication. Phosphorylated by ATM in response to ionizing irradiation. ATM and ATR can both phosphorylate Ser-317 and Ser-345 and this results in enhanced kinase activity. Phosphorylation at Ser-345 also increases binding to 14-3-3 proteins and promotes nuclear retention. Conversely, dephosphorylation at Ser-345 by PPM1D may contribute to exit from checkpoint mediated cell cycle arrest. May also be phosphorylated at Ser-280 by AKT1/PKB, which may promote mono and/or diubiquitination. Also phosphorylated at undefined residues during mitotic arrest, which results in decreased activity. Ref.1 Ref.9 Ref.13 Ref.16 Ref.17 Ref.20 Ref.27 Ref.36 Ref.21 Ref.30 Ref.32 Ref.28 Ref.8 Ref.10 Ref.19 Ref.26 Ref.38 Ref.39 Ref.40
Ubiquitinated. Mono or diubiquitination promotes nuclear exclusion By similarity.
SIMILARITY:Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. NIM1 subfamily.
Contains 1 protein kinase domain.
Product Usage Statements
Quality Assurance
Routinely evaluated by Western blot on K562 whole cell lysate.
Western Blot Analysis:
A 1:1000 dilution of this lot was used to detect Chk1 K562 whole cell lysate
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
DNA damage activates ATM, which phosphorylates and activates CHK1 and CHK2/Cds1. The principal substrates of CHK1 and CHK2 are cdc25C (inhibited by phosphorylation), p53 (stabilized by phosphorylation), and BRCA1 (scatters through the nucleus following phosphorylation). Activation of p53 results in cell cycle arrest, or induction of apoptosis. Cdc25C inhibition prevents dephosphorylation of the inhibitory phosphotyrosine residue on cdc2/cdk1, thus preventing the G2/M transition. BRCA1 dispersion through the nucleus is associated with DNA repair.
Product Information
Format
Culture Supernatant
Control
K562 whole cell lysate.
Presentation
Unpurified rabbit monoclonal IgG in buffer containing 50 mM Tris-Glycine (pH 7.4), 0.15 M NaCl, 40% Glycerol, 0.01% sodium azide and 0.05% BSA
Applications
Application
Please note that this product will not be available for sale after March 15, 2015. Please select one of the other antibodies against this target. This Anti-Chk1 (N-term) Antibody, clone E250, Rabbit is validated for use in WB, IC for the detection of Chk1 (N-term).
Key Applications
Immunocytochemistry
Western Blotting
Application Notes
Immunocytochemistry:
A previous lot of this antibody was used on Immunocytochemistry
Biological Information
Immunogen
Synthetic peptide directed against the N-terminus of human Chk1.
Epitope
N-Terminus
Clone
E250
Host
Rabbit
Specificity
This antibody recognizes Chk1 at the N-Terminus.
Isotype
IgG
Species Reactivity
Rat Human
Species Reactivity Note
Proven to react with human and mouse. Not expected to react with rat.
FUNCTION:Required for checkpoint mediated cell cycle arrest in response to DNA damage or the presence of unreplicated DNA. May also negatively regulate cell cycle progression during unperturbed cell cycles. Recognizes the substrate consensus sequence [R-X-X-S/T]. Binds to and phosphorylates CDC25A, CDC25B and CDC25C. Phosphorylation of CDC25A at 'Ser-178' and 'Thr-507' and phosphorylation of CDC25C at 'Ser-216' creates binding sites for 14-3-3 proteins which inhibit CDC25A and CDC25C. Phosphorylation of CDC25A at 'Ser-76', 'Ser-124', 'Ser-178', 'Ser-279' and 'Ser-293' promotes proteolysis of CDC25A. Inhibition of CDC25 activity leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. Binds to and phosphorylates RAD51 at 'Thr-309', which may enhance the association of RAD51 with chromatin and promote DNA repair by homologous recombination. Binds to and phosphorylates TLK1 at 'Ser-743', which prevents the TLK1-dependent phosphorylation of the chromatin assembly factor ASF1A. This may affect chromatin assembly during S phase or DNA repair. May also phosphorylate multiple sites within the C-terminus of TP53, which promotes activation of TP53 by acetylation and enhances suppression of cellular proliferation. Ref.1 Ref.6 Ref.9 Ref.13 Ref.15 Ref.16 Ref.17 Ref.18 Ref.20 Ref.22 Ref.24 Ref.25 Ref.27 Ref.29 Ref.33 Ref.35 Ref.36
Catalytic activity ATP + a protein = ADP + a phosphoprotein.
SUBUNIT: Interacts with BRCA1, CLSPN, PPM1D, RAD51, TIMELESS, XPO1/CRM1 and YWHAZ/14-3-3 zeta. Ref.1 Ref.35 Ref.14 Ref.21 Ref.23 Ref.30 Ref.32 Ref.34
SUBCELLULAR LOCATION:Nucleus. Cytoplasm. Note= Nuclear export is mediated at least in part by XPO1/CRM1. Also localizes to the centrosome specifically during interphase, where it may protect centrosomal CDC2 kinase from inappropriate activation by cytoplasmic CDC25B. Ref.1 Ref.29 Ref.14 Ref.21 Ref.31
SPECIFICITY:Expressed ubiquitously with the most abundant expression in thymus, testis, small intestine and colon. Ref.1
DOMAIN: The autoinhibitory region (AIR) inhibits the activity of the kinase domain. Ref.28
PTM:Phosphorylated by ATR in a RAD17-dependent manner in response to ultraviolet irradiation and inhibition of DNA replication. Phosphorylated by ATM in response to ionizing irradiation. ATM and ATR can both phosphorylate Ser-317 and Ser-345 and this results in enhanced kinase activity. Phosphorylation at Ser-345 also increases binding to 14-3-3 proteins and promotes nuclear retention. Conversely, dephosphorylation at Ser-345 by PPM1D may contribute to exit from checkpoint mediated cell cycle arrest. May also be phosphorylated at Ser-280 by AKT1/PKB, which may promote mono and/or diubiquitination. Also phosphorylated at undefined residues during mitotic arrest, which results in decreased activity. Ref.1 Ref.9 Ref.13 Ref.16 Ref.17 Ref.20 Ref.27 Ref.36 Ref.21 Ref.30 Ref.32 Ref.28 Ref.8 Ref.10 Ref.19 Ref.26 Ref.38 Ref.39 Ref.40
Ubiquitinated. Mono or diubiquitination promotes nuclear exclusion By similarity.
SIMILARITY:Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. NIM1 subfamily.
Contains 1 protein kinase domain.
Product Usage Statements
Quality Assurance
Routinely evaluated by Western blot on K562 whole cell lysate.
Western Blot Analysis:
A 1:1000 dilution of this lot was used to detect Chk1 K562 whole cell lysate
Usage Statement
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Storage and Shipping Information
Storage Conditions
Stable for 1 year at -20ºC from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol containing solutions to become frozen during storage.
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