Product Pathways - MAPK Signaling
Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb #12280
|W IP IF-IC||H M R Mk (Z) (B) (GP)||Endogenous||38 kDa MKK6, 40 kDa MKK3||Rabbit IgG|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Z=Zebrafish B=Bovine GP=Guinea Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb recognizes endogenous levels of MKK3 and MKK6 proteins only when phosphorylated at Ser189 (MKK3) or Ser207 (MKK6).
Source / Purification
Monoclonal antibody is produced by immunizing animals with synthetic peptides corresponding to residues surrounding Ser189 of human MKK3 protein and Ser207 of human MKK6 protein.
Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated (-) or treated with TPA #4174 (200 nM, 15 min; +), using Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb (upper), MKK3 (D4C3) Rabbit mAb #8535 (middle), or MKK6 (D31D1) Rabbit mAb #8550 (lower).
Immunoprecipitation of phospho-MKK3 (Ser189)/MKK6 (Ser207) from HeLa cells, untreated or treated with TPA #4174 (200 nM, 15 min), using Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb (lanes 3 and 4) or Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lanes 5 and 6). Lanes 1 and 2 are 10% input. Western blot analysis was performed usingPhospho-MKK3 (Ser189)/MKK6 (Ser207) Antibody #9231. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or UV-treated (40 mJ/cm2 with 30 min recovery; right), using Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
MKK3 and MKK6 are two closely related dual-specificity protein kinases that activate p38 MAP kinase (1-5). MKK3 and MKK6 both phosphorylate and activate p38 MAP kinase at its activation site, Thr-Gly-Tyr, but do not phosphorylate or activate Erk1/2 or SAPK/JNK. Phosphorylation of p38 MAP kinase dramatically stimulates its ability to phosphorylate protein substrates such as ATF-2 and Elk-1. MKK3 and MKK6 are both activated by different forms of cellular stress and inflammatory cytokines (4,5). Activation of MKK3 and MKK6 occurs through phosphorylation at Ser189 and Thr222 on MKK3 (2) and Ser207 and Thr211 on MKK6 (4,5).
- Derijard, B. et al. (1995) Science 267, 682-685.
- Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
- Sluss, H.K. et al. (1994) Mol. Cell. Biol. 14, 8376-8384.
- Raingeaud, J. et al. (1996) Mol. Cell. Biol. 16(3), 1247-1255.
- Han, J. et al. (1996) J. Biol. Chem. 271, 2886-2891.
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- 8535 MKK3 (D4C3) Rabbit mAb
- 8550 MKK6 (D31D1) Rabbit mAb
- 9236 Phospho-MKK3 (Ser189)/MKK6 (Ser207) (22A8) Rabbit mAb
- 9231 Phospho-MKK3 (Ser189)/MKK6 (Ser207) Antibody
- 9230 PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit
- 9264 MKK6 Antibody
- 5674 MKK3 Antibody
- 9238 MKK3b Antibody
- 9233 MKK3/MKK6 Control Cell Extracts
- 6294 SignalSilence® MKK3 siRNA I
- 6295 SignalSilence® MKK3 siRNA II
- 4174 TPA (12-O-Tetradecanoylphorbol-13-Acetate)
- 3678 Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb
- 3900 Rabbit (DA1E) mAb IgG XP® Isotype Control
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7075 Anti-biotin, HRP-linked Antibody
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 9997 Tris Buffered Saline with Tween 20 (TBST-10X)
- 9998 BSA
- 9999 Nonfat Dry Milk
For Research Use Only. Not For Use In Diagnostic Procedures.