Revision 4
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, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

18

Source/Isotype:

Rabbit IgG

UniProt ID:

#P24844

Entrez-Gene Id:

10398

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunofluorescence (Immunocytochemistry) 1:400 - 1:800

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.

Specificity / Sensitivity

Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb recognizes endogenous levels of myosin light chain 2 protein only when phosphorylated at Thr18 and Ser19.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Thr18/Ser19 of human myosin light chain 2 protein.

Background

Myosin is composed of six polypeptide chains: two identical heavy chains and two pairs of light chains. Myosin light chain 2 (MLC2), also known as myosin regulatory light chain (MRLC), RLC, or LC20, has many isoforms depending on its distribution. In smooth muscle, MLC2 is phosphorylated at Thr18 and Ser19 by myosin light chain kinase (MLCK) in a Ca2+/calmodulin-dependent manner (1). This phosphorylation is correlated with myosin ATPase activity and smooth muscle contraction (2). ROCK also phosphorylates Ser19 of smooth muscle MLC2, which regulates the assembly of stress fibers (3). Phosphorylation of smooth muscle MLC2 at Ser1/Ser2 and Ser9 by PKC and cdc2 has been reported to inhibit myosin ATPase activity (4,5). Phosphorylation by cdc2 controls the timing of cytokinesis (5). Transgenic mice lacking phosphorylation sites on the cardiac muscle isoform show morphological and functional abnormalities (6).

  1. Ikebe, M. and Hartshorne, D.J. (1985) J. Biol. Chem. 260, 10027-10031.
  2. Tan, J. L. et al. (1992) Annu. Rev. Biochem. 61, 721-759.
  3. Totsukawa, G. et al. (2000) J. Cell Biol. 150, 797-806.
  4. Ikebe, M. et al. (2000) J. Biol. Chem. 262, 9569-9573.
  5. Satterwhite, L. L. et al. (1992) J. Cell Biol. 118, 595-605.
  6. Sanbe, A. et al. (1999) J. Biol. Chem. 274, 21085-21094.

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 IF-IC: Immunofluorescence (Immunocytochemistry)

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 4
#95777

Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb

Western Blotting Image 1: Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb Expand Image
使用 Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图),对溶剂处理 (-) 或已经 myosin 激酶抑制剂 ML-7(75 uM,30 分钟;+)处理的 HeLa 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb Expand Image
使用 Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb(上图)或 Myosin Light Chain 2 (D18E2) Rabbit mAb #8505(下图)对不同细胞系的提取物进行蛋白质印迹分析。在封闭之前,膜未经处理 (-) 或使用小牛肠磷酸酶 (CIP) 处理 (250 单位/Ml,2 人力资源;+) 在 37ºc 下。
Immunofluorescence Image 1: Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb Expand Image
使用 Phospho-Myosin Light Chain 2 (Thr18/Ser19) (E2J8F) Rabbit mAb (绿色),对未经处理 (左图) 或 λ 磷酸酶处理后 (右图) 的 Hela 细胞进行共聚焦免疫荧光分析。肌动蛋白微丝由 DyLight 554 Phalloidin #13054(红色)标记。样本用 ProLong® Gold Antifade Reagent with DAPI #8961(蓝色)封片。