Western blot analysis was performed on whole cell extracts (30 ug lysate) of HEL 92.1.7 (Lane 1), and HEK-293 (Lane 2). The blots were probed with Anti-Chk2 Mouse Monoclonal Antibody (Product# MA511940, 1-3 ug/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Secondary Antibody, HRP conjugate (Product # 626520, 1:4000 dilution). A 60 kDa band corresponding to Chk2 was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||Recombinant Chk2 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||Assay Dependent|
|Immunoprecipitation (IP)||2 µg/ mg protein lystate|
|Western Blot (WB)||1-3 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-11940 targets Chk2 in immunofluorescence, immunoprecipiation, and Western blot applications and shows reactivity with Human samples. The antibody does not react with Chk1.
The MA5-11940 immunogen is recombinant Chk2 protein.
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 functionas essential components in the G2 DNA damage checkpoint by phophorylating 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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Ionizing radiation-dependent and independent phosphorylation of the 32-kDa subunit of replication protein A during mitosis.
MA5-11940 was used in western blot to study ionizing radiation-dependent phosphorylation of the 32-kDa subunit of replication protein A during mitosis
|Stephan H,Concannon C,Kremmer E,Carty MP,Nasheuer HP||Nucleic acids research (37:6028)||2009|
Tumor suppressor protein C53 antagonizes checkpoint kinases to promote cyclin-dependent kinase 1 activation.
MA5-11940 was used in western blot to examine the role of tumor suppressor protein C53 in cyclin-dependent kinase 1 activation
|Jiang H,Wu J,He C,Yang W,Li H||Cell research (19:458)||2009|
CHK2-decreased protein expression and infrequent genetic alterations mainly occur in aggressive types of non-Hodgkin lymphomas.
MA5-11940 was used in western blot to study the potential role of CHK2 alterations in the pathogenesis of lymphoid neoplasms
|Tort F,Hernàndez S,Beà S,Martínez A,Esteller M,Herman JG,Puig X,Camacho E,Sánchez M,Nayach I,Lopez-Guillermo A,Fernández PL,Colomer D,Hernàndez L,Campo E||Blood (100:4602)||2002|
Polo-like kinase 1 and Chk2 interact and co-localize to centrosomes and the midbody.
MA5-11940 was used in immunocytochemistry and western blot to study the role of Polo-like kinase 1 and Chk2 in DNA damage checkpoint pathways
|Tsvetkov L,Xu X,Li J,Stern DF||The Journal of biological chemistry (278:8468)||2003|