Chk1 (Cell Cycle Checkpoint Kinase 1)

Size

£646.00

SKU:: 052858
Purity:: Purified
Storage Conditions:: -20°C
Supplier:: United States Biological
Host:: Mouse
Immunogen:: Purified recombinant protein corresponding to human Chk1.
Clone:: 2G1D5 (recomb)
Format:: Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 0.1mg/ml BSA, 150mM sodium chloride, <0.02% sodium azide, 50% glycerol.
Extra Details:: Chk1 kinase acts downstream of ATM/ATR kinase and plays an important role in DNA damage checkpoint control, embryonic development, and tumor suppression (1). Activation of Chk1 involves phosphorylation at Ser317 and Ser345 by ATM/ATR, followed by autophosphorylation of Ser296. Activation occurs in response to blocked DNA replication and certain forms of genotoxic stress (2). While phosphorylation at Ser345 serves to localize Chk1 to the nucleus following checkpoint activation (3), phosphorylation at Ser317 along with site-specific phosphorylation of PTEN allows for re-entry into the cell cycle following stalled DNA replication (4). Chk1 exerts its checkpoint mechanism on the cell cycle, in part, by regulating the cdc25 family of phosphatases. Chk1 phosphorylation of cdc25A targets it for proteolysis and inhibits its activity through 14-3-3 binding (5). Activated Chk1 can inactivate cdc25C via phosphorylation at Ser216, blocking the activation of cdc2 and transition into mitosis (6). Centrosomal Chk1 has been shown to phosphorylate cdc25B and inhibit its activation of CDK1-cyclin B1, thereby abrogating mitotic spindle formation and chromatin condensation (7). Furthermore, Chk1 plays a role in spindle checkpoint function through regulation of aurora B and BubR1 (8). Research studies have implicated Chk1 as a drug target for cancer therapy as its inhibition leads to cell death in many cancer cell lines (9). Applications: Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry/Immunofluorescence. Other applications not tested. Recommended Dilution: Western Blot: 1:1000, incubate membrane with diluted primary antibody in 5% nonfat dry milk, TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight. Immunofluorescence (IC): 1:200 -1:400 Immunohistochemistry (paraffin): 1:50-1:200 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Shipping Conditions:: Blue Ice