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PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit #9230
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).Learn more about how we get our images
Western blot analysis of extracts from various cell lines using MKK3 (D4C3) Rabbit mAb.Learn more about how we get our images
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.Learn more about how we get our images
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 using
Phospho-MKK3 (Ser189)/MKK6 (Ser207) Antibody #9231. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.Learn more about how we get our images
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).Learn more about how we get our images
Gallery: PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit #9230
|Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb 12280||100 µl||
||H M R Mk||38 kDa MKK6, 40 kDa MKK3||Rabbit IgG|
|MKK3 (D4C3) Rabbit mAb 8535||100 µl||
||H M R Mk||38-40||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
|Anti-biotin, HRP-linked Antibody 7075||100 µl||
|20X LumiGLO® Reagent and 20X Peroxide 7003||5 ml each||
|Biotinylated Protein Ladder Detection Pack 7727||100 µl||
|MKK3/MKK6 Control Cell Extracts 9233||200 µl||
The PhosphoPlus® MKK3/MKK6 (Ser189/207) Antibody Kit provides reagents and protocols for the rapid analysis of the phosphorylation status of MKK3/MKK6 at Ser189/207.
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). MKK3 (D4C3) Rabbit mAb recognizes endogenous levels of total MKK3 protein. This antibody cross-reacts slightly with MKK6 but demonstrates an approximate 10-fold preference for MKK3 using recombinant, purified proteins.
Monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser189 of human MKK3 protein and Ser207 of human MKK6 protein and with a synthetic peptide corresponding to residues near the carboxy terminus of human MKK3 protein.
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).
For Research Use Only. Not For Use In Diagnostic Procedures. Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc. PhosphoPlus® is a trademark of Cell Signaling Technology, Inc. LumiGLO® is a registered trademark of Kirkegaard & Perry Laboratories.