Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-SAPK/JNK Pathway Antibody Sampler Kit #9912

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668 40 µl W IP IHC-P H M R Dm Sc 46, 54 Rabbit IgG
Phospho-SEK1/MKK4 (Ser257) (C36C11) Rabbit mAb #4514 40 µl W F H M R Mk 44 Rabbit
Phospho-ATF-2 (Thr71) (11G2) Rabbit mAb #5112 40 µl W F H M R Mk 70 Rabbit IgG
Phospho-c-Jun (Ser63) (54B3) Rabbit mAb #2361 40 µl W IHC-P H M R 48 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  Dm=D. melanogaster  Sc=S. cerevisiae
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each antibody in the Phospho-SAPK/JNK Antibody Sampler Kit recognizes endogenous levels of the phosphorylated form of its specific target.

Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or anisomycin-treated C6 cells, or untreated or UV-treated 293 cells, using Phospho-c-Jun (Ser63) (54B3) Rabbit mAb #2361 (upper) or c-Jun (60A8) Rabbit mAb #9165 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated or treated with sorbitol or UV, as indicated, using Phospho-SEK1/MKK4 (Ser257) (C36C11) Rabbit mAb #4514.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells, untreated or UV-treated, NIH/3T3 cells, untreated or UV-treated and C6 cells, untreated or anisomycin-treated, using Phospho-SAPK/JNK (Thr183/Tyr185) (81E11) Rabbit mAb #4668.


Western Blotting

Western Blotting

Western blot analysis of extracts from untreated or UV-treated COS cells, NIH/3T3 cells and C6 cells, using Phospho-ATF-2 (Thr71) (11G2) Rabbit mAb #5112 (upper), or ATF-2 (20F1) Rabbit mAb #9226 (lower).

Description

The Phospho-SAPK/JNK Pathway Antibody Sampler Kit provides a fast and economical means of evaluating multiple members of the SAPK/JNK pathway as well as their activation state. The kit contains enough primary and secondary antibodies to perform four Western blot experiments.

Source / Purification

Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptides corresponding to residues surrounding Ser257 of human SEK1, Thr183/Tyr185 of human SAPK, Thr71 of human ATF-2 or Ser63 of human c-Jun.

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).

  1. Davis, R.J. (1999) Biochem Soc Symp 64, 1-12.
  2. Ichijo, H. (1999) Oncogene 18, 6087-93.
  3. Kyriakis, J.M. and Avruch, J. (2001) Physiol Rev 81, 807-69.
  4. Kyriakis, J.M. (1999) J Biol Chem 274, 5259-62.
  5. Leppä, S. and Bohmann, D. (1999) Oncogene 18, 6158-62.
  6. Whitmarsh, A.J. and Davis, R.J. (1998) Trends Biochem Sci 23, 481-5.

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