Preparation And Storage
Recommended Assay Procedures
G151-666 immunoprecipitates inactive JNK1 and JNK2 kinases, therefore the antibody is not recommended for in vitro kinase assays. G151-666 is most useful for detection of JNK2 or for detection of both JNK1 and JNK2 in the same assay. Clone G151-333 (Cat. No. 554286) appears to be stronger for detection of JNK1 and is generally suggested for most applications involving JNK1. Clone G151-333 can be used to immunoprecipitate an active JNK1 kinase. HeLa cells (ATCC CCL-1) or NIH-3T3 mouse embryonic fibroblasts (ATCC CRL-1658) are suggested as a positive control for western blot analysis.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
C-Jun NH2-terminal kinase (JNK) binds to the c-Jun terminal transactivation domain and phosphorylates it on Ser-63 and Ser-73. Phosphorylation enhances the transcriptional activity of c-Jun. The Ser-Pro-acidic sequence targeted by JNK kinase activity establishes it as a prolinedirected kinase related to the MAP kinases and cyclin/dependent kinases. JNK may act as a tumor promoter in response to UV-irradiation since its activity is potently stimulated by radiation. This has relevance to observations that c-Jun transcriptional activity is upregulated by UV irradiation. In addition to UV irradiation, JNK is also activated by some other forms of cellular stress, including heatshock. Both the JNK1 (46 kDa) and JNK2 (54 kDa) isozymes appear equally capable of binding to the c-Jun terminal transactivation domain following induction by UV irradiation or heatshock. G151-666 recognizes both the JNK1 and JNK2 isozymes of JNK1. A bacterially expressed fusion protein of human JNK1 was used as immunogen.
Development References (6)
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Adler V, Fuchs SY, Kim J. jun-NH2-terminal kinase activation mediated by UV-induced DNA lesions in melanoma and fibroblast cells. Cell Growth Differ. 1995; 6(11):1437-1446. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Adler V, Pincus MR, Brandt-Rauf PW, Ronai Z. Complexes of p21RAS with JUN N-terminal kinase and JUN proteins. Proc Natl Acad Sci U S A. 1995; 92(23):10585-10589. (Clone-specific: Immunoprecipitation). View Reference
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Adler V, Schaffer A, Kim J, Dolan L, Ronai Z. UV irradiation and heat shock mediate JNK activation via alternate pathways. J Biol Chem. 1995; 270(44):26071-26077. (Clone-specific: Immunoprecipitation, Western blot). View Reference
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Devary Y, Rosette C, DiDonato JA, Karin M. NF-kappa B activation by ultraviolet light not dependent on a nuclear signal. Science. 1993; 261(5127):1442-1445. (Biology). View Reference
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Dérijard B, Hibi M, Wu IH. JNK1: a protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain. Cell. 1994; 76(6):1025-1037. (Biology). View Reference
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Hibi M, Lin A, Smeal T, Minden A, Karin M. Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. Genes Dev. 1993; 7(11):2135-2148. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.