Phospho-Chk1 (S296) Recombinant Rabbit Monoclonal Antibody [SN06-50] (ET1611-76)
Catalog# ET1611-76
Rabbit Monoclonal to Phospho-Chk1 (S296)
-
WB
-
IHC-P
-
IF-Cell
-
Human
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Mouse
-
Rat
Western blot analysis of Phospho-Chk1 (S296) on different lysates with Rabbit anti-Phospho-Chk1 (S296) antibody (ET1611-76) at 1/1,000 dilution.
Lane 1: HEK-293 cell lysate
Lane 2: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate
Lane 3: HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, then the membrane treated with λpp for 1 hour
Lane 4: NIH/3T3 cell lysate
Lane 5: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate
Lane 6: NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, then the membrane treated with λpp for 1 hour
Lane 7: C6 cell lysate
Lane 8: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate
Lane 9: C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: Lane 1-3: 20 seconds; Lane 4-9: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-76) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
Specifications
Product Type
Recombinant Rabbit monoclonal primary
Product Name
Phospho-Chk1 (S296) Recombinant Rabbit Monoclonal Antibody [SN06-50] (ET1611-76)
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding ser296 of human chk1
Host
Rabbit
Modification
Phospho
Modification Site
S296
Positive Control
HEK-293 treated with 200nM Calyculin A for 1 hour cell lysate, NIH/3T3 treated with 100nM Calyculin A for 30 minutes cell lysate, C6 treated with 100ng/mL Calyculin A for 1 hour cell lysate, human tonsil tissue, human breast cancer tissue, HEK-293 cells treated with 100nM Calyculin A for 30 minutes.
Conjugation
Unconjugated
Clonality
Monoclonal
Clone Number
SN06-50
RRID
APPLICATION DILUTION
-
WB
-
1:1,000
-
IHC-P
-
1:200-1:1,000
-
IF-Cell
-
1:100
PROPERTIES
Form
Liquid
Storage Condition
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration
1ug/ul
Purification
Protein A affinity purified.
Molecular Weight
Predicted band size: 54 kDa
Isotype
IgG
TARGET
UNIPROT #
PROTEIN NAME
Phospho-Chk1 (S296)
SYNONYMS
C85740 antibody;Cell cycle checkpoint kinase antibody;Checkpoint , S. pombe, homolog of, 1 antibody;Checkpoint kinase 1 antibody;Checkpoint kinase 1 homolog (S. pombe) antibody;CHEK 1 antibody;Chek1 antibody;Chk 1 antibody;Chk1 antibody;CHK1 checkpoint homolog (S. pombe) antibody;CHK1_HUMAN antibody;EC 2.7.11.1 antibody;rad27 antibody;Serine/threonine protein kinase Chk1 antibody;Serine/threonine-protein kinase CHK1 antibody;STT3, subunit of the oligosaccharyltransferase complex, homolog A (S. cerevisiae) antibody
SEQUENCE SIMILARITIES
Belongs to the protein kinase superfamily. CAMK Ser/Thr protein kinase family. NIM1 subfamily.
TISSUE SPECIFICITY
Expressed ubiquitously with the most abundant expression in thymus, testis, small intestine and colon.
POST-TRANSLATIONAL MODIFICATION
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 (By similarity). 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.
SUBCELLULAR LOCATION
Chromosome, Cytoplasm, Cytoskeleton, Nucleus.
FUNCTION
Cell cycle events are regulated by the sequential activation and deactivation of cyclin dependent kinases (Cdks) and by proteolysis of cyclins. Chk1 and Chk2 are involved in these processes as regulators of Cdks. Chk1 and Chk2 both function as essential components in the G2 DNA damage checkpoint by phosphorylating Cdc25C in response to DNA damage. Phosphorylation inhibits Cdc25C activity, thereby blocking mitosis. Cdc25A, Cdc25B and Cdc25C protein tyrosine phosphatases function as mitotic activators by dephosphorylating Cdc2 p34 on regulatory tyrosine residues. It has also been shown that Chk1 can phosphorylate Wee1 in vitro, providing evidence that the hyperphosphorylated form of Wee1, seen in cells delayed by Chk1 overexpression, is due to phosphorylation by Chk1.
