• Western blot analysis of extracts of various cell lines, using Phospho-Chk1-S345 antibody (STJ29344). Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% BSA.
  • Western blot analysis of NIH/3T3, using Phospho-Chk1-S345 Rabbit polyclonal antibody (STJ29344) at 1:500 dilution. NIH/3T3 cells were treated by UV at room temperature for 15-30 minutes. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) (STJS000856) at 1:10000 dilution. Lysates/proteins: 25 Mu g per lane. Blocking buffer: 3% nonfat dry milk in TBST. Detection: ECL Basic Kit. Exposure time: 180s.
  • Western blot analysis of extracts of C6 cells, using Phospho-Chk1-S345 polyclonal antibody (STJ29344) at 1:1000 dilution or Chk1 antibody (A7653). C6 cells were treated by ATP (5 mM) at 30 °C for 1 hour. Secondary antibody: HRP Goat Anti-rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% BSA. Detection: ECL Enhanced Kit. Exposure time: 5min.

Anti-Phospho-CHEK1-S345 antibody (STJ29344)

SKU:
STJ29344

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Host: Rabbit
Applications: WB
Reactivity: Human/Mouse/Rat
Note: STRICTLY FOR FURTHER SCIENTIFIC RESEARCH USE ONLY (RUO). MUST NOT TO BE USED IN DIAGNOSTIC OR THERAPEUTIC APPLICATIONS.
Short Description: Rabbit polyclonal antibody anti-Phospho-Chk1-S345 is suitable for use in Western Blot research applications.
Clonality: Polyclonal
Conjugation: Unconjugated
Isotype: IgG
Formulation: PBS with 0.01% Thimerosal, 50% Glycerol, pH7.3.
Purification: Affinity purification
Dilution Range: WB 1:100-1:500
Storage Instruction: Store at-20°C for up to 1 year from the date of receipt, and avoid repeat freeze-thaw cycles.
Gene Symbol: CHEK1
Gene ID: 1111
Uniprot ID: CHK1_HUMAN
Immunogen: A synthetic phosphorylated peptide around S345 of human Chk1 (NP_001265.2).
Immunogen Sequence: SFSQP
Tissue Specificity Expressed ubiquitously with the most abundant expression in thymus, testis, small intestine and colon.
Post Translational Modifications 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 induces a change in the conformation of the protein, activates the kinase activity and is a prerequisite for interaction with FBXO6 and subsequent ubiquitination at Lys-436. 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. Phosphorylation at Ser-280 by AKT1/PKB, may promote mono and/or diubiquitination. Also phosphorylated at undefined residues during mitotic arrest, resulting in decreased activity. Ubiquitinated. Mono or diubiquitination promotes nuclear exclusion. The activated form (phosphorylated on Ser-345) is polyubiquitinated at Lys-436 by some SCF-type E3 ubiquitin ligase complex containing FBXO6 promoting its degradation. Ubiquitination and degradation are required to terminate the checkpoint and ensure that activated CHEK1 does not accumulate as cells progress through S phase, when replication forks encounter transient impediments during normal DNA replication. 'Lys-63'-mediated ubiquitination by TRAF4 at Lys-132 activates cell cycle arrest and activation of DNA repair. Proteolytically cleaved at the C-terminus by SPRTN during normal DNA replication, thereby promoting CHEK1 removal from chromatin and activating the protein kinase activity.
Function Serine/threonine-protein kinase which is required for checkpoint-mediated cell cycle arrest and activation of DNA repair in response to the presence of DNA damage or unreplicated DNA. May also negatively regulate cell cycle progression during unperturbed cell cycles. This regulation is achieved by a number of mechanisms that together help to preserve the integrity of the genome. 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. Phosphorylation of CDC25A at 'Ser-76' primes the protein for subsequent phosphorylation at 'Ser-79', 'Ser-82' and 'Ser-88' by NEK11, which is required for polyubiquitination and degradation of CDCD25A. Inhibition of CDC25 leads to increased inhibitory tyrosine phosphorylation of CDK-cyclin complexes and blocks cell cycle progression. Also phosphorylates NEK6. Binds to and phosphorylates RAD51 at 'Thr-309', which promotes the release of RAD51 from BRCA2 and enhances the association of RAD51 with chromatin, thereby promoting DNA repair by homologous recombination. Phosphorylates multiple sites within the C-terminus of TP53, which promotes activation of TP53 by acetylation and promotes cell cycle arrest and suppression of cellular proliferation. Also promotes repair of DNA cross-links through phosphorylation of FANCE. Binds to and phosphorylates TLK1 at 'Ser-743', which prevents the TLK1-dependent phosphorylation of the chromatin assembly factor ASF1A. This may enhance chromatin assembly both in the presence or absence of DNA damage. May also play a role in replication fork maintenance through regulation of PCNA. May regulate the transcription of genes that regulate cell-cycle progression through the phosphorylation of histones. Phosphorylates histone H3.1 (to form H3T11ph), which leads to epigenetic inhibition of a subset of genes. May also phosphorylate RB1 to promote its interaction with the E2F family of transcription factors and subsequent cell cycle arrest. Phosphorylates SPRTN, promoting SPRTN recruitment to chromatin. Reduces replication stress and activates the G2/M checkpoint, by phosphorylating and inactivating PABIR1/FAM122A and promoting the serine/threonine-protein phosphatase 2A-mediated dephosphorylation and stabilization of WEE1 levels and activity. Isoform 2: Endogenous repressor of isoform 1, interacts with, and antagonizes CHK1 to promote the S to G2/M phase transition.
Protein Name Serine/Threonine-Protein Kinase Chk1
Chk1 Checkpoint Homolog
Cell Cycle Checkpoint Kinase
Checkpoint Kinase-1
Database Links Reactome: R-HSA-1433557
Reactome: R-HSA-176187
Reactome: R-HSA-5693607
Reactome: R-HSA-5693616
Reactome: R-HSA-6796648
Reactome: R-HSA-6804756
Reactome: R-HSA-69473
Reactome: R-HSA-69601
Reactome: R-HSA-75035
Reactome: R-HSA-8953750
Cellular Localisation Nucleus
Chromosome
Cytoplasm
Cytoskeleton
Microtubule Organizing Center
Centrosome
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
Proteolytic Cleavage At The C-Terminus By Sprtn Promotes Removal From Chromatin
Alternative Antibody Names Anti-Serine/Threonine-Protein Kinase Chk1 antibody
Anti-Chk1 Checkpoint Homolog antibody
Anti-Cell Cycle Checkpoint Kinase antibody
Anti-Checkpoint Kinase-1 antibody
Anti-CHEK1 antibody
Anti-CHK1 antibody

Information sourced from Uniprot.org

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