Technical Data
S0096-04
SAPK1, JNK (Stress Activated Protein Kinase, Jun Kinase)
Description:
Stress-activated protein (SAP) kinases are proline-directed, serine/threonine-specific kinases that require phosphorylation on tyrosine for phosphotransferase activity.SAP Kinase was originally shown to phosphorylate microtubule-associated protein-2, but c-Jun appears to be an important physiological target of this kinase. Consequently SAPK has also been called Jun Kinase.3 Phosphorylation of Ser-63 and Ser-73 of c-Jun by SAP kinases promotes activation of the transcription factor AP-1. cDNA cloning studies with rat and human libraries have shown that there are several isoforms of the SAP Kinase family. Many stimuli activate SAP Kinase, including heat shock, 7 hyperosmotic shock, UV irradiation, exposure to protein synthesis inhibitors such as cycloheximide and anisomycin, and treatment with proinflammatory cytokines such as tumor necrosis factor-a.

Applications:
Suitable for use in Immunoblot anaylsis and Immunoprecipitation. Other applications have not been tested.

Recommended Dilution:
Immunoblot Analysis: 1ug/ml detected SAPK1 in RIPA lysates from human A431, murine 3T3 and rat PC-12 cells. At 1ug/ml, the signal detected with 10ng of gst-SAPK1a was approximately three times greater than the signal detected of with 10ng gst-SAPK1b.

Included Positive Antigen Control: Non-stimulated A431 cell lysate. Use 20ug per lane for minigels.

Immunoprecipitation Protein Kinase Assay: 4ug immunoprecipitated active recombinant rat SAPK1a, which then catalyzed the transfer of the y-phosphate of [y32 P]-ATP to c-Jun protein substrate. Immunoblot Analysis A431 cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with anti-SAPK1 (1ug/ml). Proteins were visualized using a goat anti-rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system.

Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4C for short-term only. For long-term storage, store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
TypeIsotypeCloneGrade
PabIgGAffinity Purified
SizeStorageShippingSourceHost
200ug4C (-20C Glycerol)Blue IceRatRabbit
Concentration:
~1mg/ml
Immunogen:
GST fusion protein corresponding to full-length rat SAPK1a. This sequence is approximately 85% identical to rat SAPK1b and rat SAPK1c. These sequences are highly conserved among mammalian species.
Purity:
Purified by Protein A affinity chromatography.
Form
Supplied as a liquid in 0.01M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol.
Specificity:
Recognizes SAPK1a (54kD). Weakly recognizes SAPK1b (54kD). The protein detected with a MWapp of 45kD may be SAPK1c. Species Reactivity: Human, mouse and rat.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.
General 1. Mukhopadhyay, N.K., et al., J. Biol. Chem 267: 33253335, 1992. 2. Kyriakis, J.M., et al., J. Biol. Chem. 266: 1004310046, 1991. 3. Davis, R.J., Trends Biochem. Sci. 19: 470475, 1994. 4. Hibi, M., et al., Genes Dev. 7: 21352148, 1993. 5. Smeal, T., et al., EMBO J. 13: 60066010, 1994. 6. Derijard, B., et al., Cell 76: 10251030, 1994. 7. Kyriakis, J.M., et al., Nature 369: 156160, 1994. 8. Sluss, H.K., et al., Mol. Cell. Biol. 14: 83768384, 1994. 9. Sanchez, et al., Nature 372: 794798, 1994. 10. Kyriakis, J.M., and Avruch, J., J. Biol. Chem 265: 1735517363, 1990.