| 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. |
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| Catalog # | S0096-04 |
| 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 [y–32 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 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. 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. |
| CAS Number | n/a |
| Clone Type | Polyclonal |
| Isotype | IgG |
| Host | Rabbit |
| Source | Rat |
| Concentration | ~1mg/ml |
| Form | Supplied as a liquid in 0.01M Tris-glycine, pH 7.4, 0.15M sodium chloride, 0.05% sodium azide, 30% glycerol. |
| Purity | Purified by Protein A affinity chromatography. |
| 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. |
| 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. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
