Cell Signaling Technology

Product Pathways - MAPK Signaling

Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb #12280

No. Size Price
12280S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价 防伪查询
12280 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 38 kDa MKK6, 40 kDa MKK3 Rabbit IgG
IP 1:100
IF-IC 1:400

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry),


Species predicted to react based on 100% sequence homology: Rat, Monkey, Zebrafish, Bovine, Guinea Pig,

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).

Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9)Rabbit mAb兔单抗能够识别内源性Ser189 (MKK3)或Ser207 (MKK6)位点被磷酸化的MKK3和MKK6蛋白。

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.




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).未处理(左)或UV-处理40 mJ/cm2 with 30 min recovery;右)的HeLa细胞,使用Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb(绿色)进行激光共聚焦免疫荧光分析。肌动蛋白丝使用DY-554鬼笔环肽标记(红色)。蓝色假色= DRAQ5® #4084 (fluorescent DNA dye).

Western Blotting

Western Blotting

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).未处理(-)或使用TPA #4174 (200 nM, 15 min; +)处理的HeLa和NIH/3T3细胞提取物,使用Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9)Rabbit mAB(上),MKK3 (D4C3)Rabbit mAb #8535(中)或MKK6 (D31D1)Rabbit mAb #8550(下)进行western blot分析。



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.未处理或经过TPA #4174 (200 nM, 15 min)处理的HeLa细胞phospho-MKK3 (Ser189)/MKK6 (Ser207),使用Phospho-MKK3 (Ser189)/MKK6 (Ser207) (D8E9) Rabbit mAb (lanes 3 and 4) 或Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lanes 5和 6)进行免疫沉淀实验。Lane1和2是10%的上样量。使用Phospho-MKK3 (Ser189)/MKK6 (Ser207) 抗体#9231进行western blot分析。Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678可以用作二抗。


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).

MKK3和MKK6是两种密切相关的双特异性蛋白激酶,能够激活p38 MAPK(1-5)。MKK3和MKK6能够通过磷酸化p38 MAPK的活化位点Thr-Gly-Tyr从而使p38 MAPK激活,但不能磷酸化或激活Erk1/2或SAPK/JNK。p38 MAPK的磷酸化能够显著提高其磷酸化蛋白底物的能力,这些底物包括ATF-2和Elk-1。MKK3和MKK6能被不同形式的细胞压力和炎症细胞因子激活(4,5)。通过磷酸化MKK3的Ser189和Thr222位点(2)以及MKK6的Ser207和Thr211位点(4,5)可以激活MKK3和MKK6。

  1. Derijard, B. et al. (1995) Science 267, 682-685.
  2. Raingeaud, J. et al. (1995) J. Biol. Chem. 270, 7420-7426.
  3. Sluss, H.K. et al. (1994) Mol. Cell. Biol. 14, 8376-8384.
  4. Raingeaud, J. et al. (1996) Mol. Cell. Biol. 16(3), 1247-1255.
  5. Han, J. et al. (1996) J. Biol. Chem. 271, 2886-2891.

Application References

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