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Phospho-CHK2 (Thr68) Mouse anti-Human, Mouse, PE, Clone: ebchk2, eBioscience
Mouse Monoclonal Antibody
Manufacturer: Invitrogen 12950841
Description: This ebchk2 monoclonal antibody recognizes human and mouse Checkpoint Kinase 2 (Chk2) when phosphorylated on threonine 68 (T68). Chk2 is a cell cycle checkpoint regulator and tumor suppressor. Along with Chk1, it plays a critical role in the cellular response to DNA damage, and is particularly important during G2/M transition. Chk2 is activated via phosphorylation on T68 by the kinase ATM, which responds primarily to double-stranded DNA breaks. Upon activation, Chk2 functions include inhibition of Cdc25 family phosphatases and activation of p53, both of which prevent progression of the cell cycle. Abnormal regulation of the ATM-Chk2 pathway is often observed in cancer. Applications Reported: This ebchk2 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This ebchk2 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.125 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Staining Protocol: All protocols work well for this monoclonal antibody.Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered. Checkpoint kinase 2 (Chk2) is a serine/threonine-protein kinase which functions as a cell cycle checkpoint regulator. ATM and ATR kinases are activated in response to ionizing and ultraviolet radition, and phosphorylate Thr68 and other sites in the amino-terminal region of Chk2 dimerisation and activation. Activated Chk2 phosphorylates and inhibits Cdc25C phosphates, preventing the entry into mitosis, and has been shown to stabilize the tumor suppressor protein p53, leading to cell cycle arrest in G1. In addition, Chk2 can phosphorylate BRCA1, allowing BRCA1 to restore survival after DNA damage. Chk2 is a putative tumor suppressor protein, with mutations in to the Chk2 gene linked to Li-Fraumeni syndrome, a familiar cancer usually associated with inherited mutations in TP53. Also, mutations in this gene are thought to confer a predisposition to sarcomas, breast cancer, and brain tumors.
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
|CDS1, CHEK2, HuCds1, PP1425, RAD53|
|4° C, store in dark, DO NOT FREEZE!|
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For Research Use Only.