04-207 | Anti-Chk1 (NT) Antibody, clone E250, rabbit monoclonal

04-207
100 µL  
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      Overview

      Replacement Information

      Key Spec Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      H, RWB, ICCRbCulture SupernatantMonoclonal Antibody
      Description
      Catalogue Number04-207
      Replaces04-207
      Brand Family Upstate
      Trade Name
      • Upstate
      DescriptionAnti-Chk1 (NT) Antibody, clone E250, rabbit monoclonal
      Alternate Names
      • CHK1 (checkpoint, S.pombe) homolog
      • CHK1 checkpoint homolog
      • CHK1 checkpoint homolog (S. pombe)
      • Checkpoint, S. pombe, homolog of, 1
      Background InformationDNA 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.
      References
      Product Information
      FormatCulture Supernatant
      Control
      • K562 whole cell lysate.
      PresentationUnpurified 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
      ApplicationPlease 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
      • Western Blotting
      • Immunocytochemistry
      Application NotesImmunocytochemistry:
      A previous lot of this antibody was used on Immunocytochemistry
      Biological Information
      ImmunogenSynthetic peptide directed against the N-terminus of human Chk1.
      EpitopeN-Terminus
      CloneE250
      HostRabbit
      SpecificityThis antibody recognizes Chk1 at the N-Terminus.
      IsotypeIgG
      Species Reactivity
      • Human
      • Rat
      Species Reactivity NoteProven to react with human and mouse. Not expected to react with rat.
      Antibody TypeMonoclonal Antibody
      Entrez Gene Number
      Gene Symbol
      • CHK1
      • EC 2.7.11.1
      Purification MethodUnpurified
      UniProt Number
      UniProt SummaryFUNCTION: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.

      Molecular Weight54 kDa
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Quality AssuranceRoutinely 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 ConditionsStable 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.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      Certificates of Analysis

      TitleLot Number
      Anti-Chk1, clone E250 - 2386942 2386942
      Anti-Chk1, clone E250 - 2386948 2386948
      Anti-Chk1, clone E250 - 2072891 2072891
      Anti-Chk1, clone E250 - 2179312 2179312
      Anti-Chk1, clone E250 - 2197860 2197860
      Anti-Chk1, clone E250 - 2251742 2251742
      Anti-Chk1, clone E250 - 2269043 2269043
      Anti-Chk1, clone E250 - 2343812 2343812
      Anti-Chk1, clone E250 - 2358805 2358805
      Anti-Chk1, clone E250 - 2364193 2364193

      References

      Reference overviewPub Med ID
      Evaluation of novel imidazotetrazine analogues designed to overcome temozolomide resistance and glioblastoma regrowth.
      Ramirez, YP; Mladek, AC; Phillips, RM; Gynther, M; Rautio, J; Ross, AH; Wheelhouse, RT; Sakaria, JN
      Molecular cancer therapeutics  14  111-9  2015

      Show Abstract
      25351918 25351918

      Brochure

      Title
      Advancing cancer research: From hallmarks & biomarkers to tumor microenvironment progression
      Product Selection Guide - Antibodies, small molecule inhibitors, kits, assays and proteins for signaling research.