Revision 1

#9127Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-IC, FC-FP

REACTIVITY:

H R Mk

SENSITIVITY:

Endogenous

MW (kDa):

45

SOURCE:

Rabbit

UniProt ID:

#Q02750

Entrez-Gene Id:

5604

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Immunocytochemistry) 1:100
Flow Cytometry (Fixed/Permeabilized) 1:50

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.

Specificity / Sensitivity

Phospho-MEK1 (Thr286) Antibody detects endogenous levels of MEK1 phosphorylated at threonine 286. This antibody does not cross-react with phosphorylated MEK2.

Species Reactivity:

Human, Rat, Monkey

Species predicted to react based on 100% sequence homology

Mouse

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Thr286 of human MEK1. Antibodies are purified by protein A and peptide affinity chromatography.

Background

MEK1 and MEK2, also called MAPK or Erk kinases, are dual-specificity protein kinases that function in a mitogen activated protein kinase cascade controlling cell growth and differentiation (1-3). Activation of MEK1 and MEK2 occurs through phosphorylation of two serine residues at positions 217 and 221, located in the activation loop of subdomain VIII, by Raf-like molecules. MEK1/2 is activated by a wide variety of growth factors and cytokines and also by membrane depolarization and calcium influx (1-4). Constitutively active forms of MEK1/2 are sufficient for the transformation of NIH/3T3 cells or the differentiation of PC-12 cells (4). MEK activates p44 and p42 MAP kinase by phosphorylating both threonine and tyrosine residues at sites located within the activation loop of kinase subdomain VIII.
MEK1 is phosphorylated at Ser298 by PAK1, which facilitates signal transduction from Raf to MEK1 and Erk2 (5-7). MEK1 is also phosphorylated by cdk5 at Thr286 in mitotic cells, causing negative feedback of the p44/42 MAP kinase pathway (8).

  1. Crews, C.M. et al. (1992) Science 258, 478-480.
  2. Alessi, D.R. et al. (1994) EMBO J. 13, 1610-19.
  3. Rosen, L.B. et al. (1994) Neuron 12, 1207-21.
  4. Cowley, S. et al. (1994) Cell 77, 841-52.
  5. Xu, B. et al. (1999) J. Biol. Chem. 274, 34029-34035.
  6. Coles, L.C. and Shaw, P.E. (2002) Oncogene 21, 2236-2244.
  7. Eblen, S. T. et al. (2002) Mol. Cell. Biol. 22, 6023-6033.
  8. Sharma, P. et al. (2002) J. Biol. Chem. 277, 528-534.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

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

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 1
#9127

Phospho-MEK1 (Thr286) Antibody

Western Blotting Image 1: Phospho-MEK1 (Thr286) Antibody Expand Image
使用 Phospho-MEK1 (Thr286) Antibody (上图) 或 MEK1 Antibody #9124(下图),对未经处理或诺考达唑处理 A431、COS 和 pC12 细胞的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Phospho-MEK1 (Thr286) Antibody Expand Image
使用 Phospho-MEK1 (Thr286) Antibody,对未经处理或 Nocodazole 处理的 COS 细胞提取物进行免疫沉淀,之后进行蛋白质印迹分析。
Immunofluorescence Image 1: Phospho-MEK1 (Thr286) Antibody Expand Image
使用 Phospho-MEK1 (Thr286) Antibody (绿色) 对分裂期 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白纤丝用 Alexa Fluor®555 phalloidin(红色)进行标记。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Phospho-MEK1 (Thr286) Antibody Expand Image
与碘化丙啶 (DNA 含量) 相比较,使用 Phospho-MEK1 (Thr286) Antibody 对未经处理的 Jurkat 细胞进行流式细胞分析。该盒显示磷酸 MEK1 阳性细胞。