Revision 6
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, IHC-P, FC-FP

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

45

Source/Isotype:

Rabbit IgG

UniProt ID:

#P36507, #Q02750

Entrez-Gene Id:

5605, 5604

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:2000
Immunohistochemistry (Paraffin) 1:50
Flow Cytometry (Fixed/Permeabilized) 1:100

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #75262.

Specificity / Sensitivity

Phospho-MEK1/2 (Ser 221) (166F8) Rabbit mAb detects endogenous levels of MEK1/2 only when phosphorylated at serine 221.

Species Reactivity:

Human, Mouse, Rat, Monkey

Species predicted to react based on 100% sequence homology

Dog

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser221 of human MEK1/2.

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.

  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.

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 IHC-P: Immunohistochemistry (Paraffin) 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.
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 6
#2338

Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb

Western Blotting Image 1: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb (上图) 和 MEK1/2 Antibody#9122 (下图),对未经处理或经 TPA 和紫外线处理的 HeLa 和 NIH/3T3 细胞提取物进行蛋白质印迹分析。
Immunohistochemistry Image 1: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser 221) (166F8) Rabbit mAb,对未经处理 (左图) 或 Raf1 激酶抑制剂 1 (右图) 处理的福尔马林固定、石蜡包埋的 A375 细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221)(166F8) Rabbit mAb,对石蜡包埋的人类乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 3: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb,对石蜡包埋的人肺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb,对未经处理 (左图) 或 λ 磷酸酶处理 (右图) 的石蜡包埋前列腺癌进行免疫组织化学分析。
Flow Cytometry Image 1: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(绿色)或用 λ-磷酸酶固定后再处理(蓝色)的血清饥饿 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。
Flow Cytometry Image 2: Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb Expand Image
使用 Phospho-MEK1/2 (Ser221) (166F8) Rabbit mAb(实线)或浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)对未经处理(蓝色)或经过 TPA(200 nM,30 分钟;绿色)处理的血清饥饿 Jurkat 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。