| Product Includes | Product # | Volume | Cap Color | Storage Temp |
|---|---|---|---|---|
| Phospho-SAPK/JNK (Thr183/Tyr185) Capture Rabbit mAb (100X) | 32090 | 400 µl |
|
+4C |
| SAPK/JNK Detection Mouse mAb (100X) | 16945 | 400 µl |
|
+4C |
| Anti-mouse IgG, HRP-linked Antibody (1000X) | 16736 | 40 µl |
|
-20C |
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Description
CST's PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Antibody Pair is offered as an economical alternative to our PathScan® Phospho-SAPK/JNK (Thr183/Tyr185) Sandwich ELISA Kit #7325. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-SAPK/JNK (Thr183/Tyr185) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by SAPK/JNK Detection Antibody and Anti-mouse IgG, HRP-conjugated Antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-SAPK/JNK (Thr183/Tyr185) protein.
Antibodies in kit are custom formulations specific to kit.
Reagents not supplied
Phosphate Buffered Saline with Tween-20 (PBST-20X) #9809
Cell Lysis Buffer (10X) #9803
TMB Substrate #7004
STOP Solution #7002
Blocking Buffer: 1X PBS/0.5% Tween-20, 1% BSA
96 Well Microplates**
Microplate Reader
** Antibody Pairs have been validated on Corning© 96 Well Clear Polystyrene High Bind Stripwell™ Microplates (#2592).
Notes: Antibody pairs have been optimized using recommended buffers, reagents, plates and the included protocol. Solutions should be made fresh daily.
Background
The stress-activated protein kinase/Jun-amino-terminal kinase SAPK/JNK is potently and preferentially activated by a variety of environmental stresses, including UV and gamma radiation, ceramides, inflammatory cytokines, and in some instances, growth factors and GPCR agonists (1-6). As with the other MAPKs, the core signaling unit is composed of a MAPKKK, typically MEKK1-MEKK4, or by one of the mixed lineage kinases (MLKs), which phosphorylate and activate MKK4/7. Upon activation, MKKs phosphorylate and activate the SAPK/JNK kinase (2). Stress signals are delivered to this cascade by small GTPases of the Rho family (Rac, Rho, cdc42) (3). Both Rac1 and cdc42 mediate the stimulation of MEKKs and MLKs (3). Alternatively, MKK4/7 can be activated in a GTPase-independent mechanism via stimulation of a germinal center kinase (GCK) family member (4). There are three SAPK/JNK genes each of which undergoes alternative splicing, resulting in numerous isoforms (3). SAPK/JNK, when active as a dimer, can translocate to the nucleus and regulate transcription through its effects on c-Jun, ATF-2, and other transcription factors (3,5).
- Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
- Ichijo, H. (1999) Oncogene 18, 6087-93.
- Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
- Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
- Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
- Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.
Background References
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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