Revision 1
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-F, IF-IC

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

80

Source/Isotype:

Rabbit IgG

UniProt ID:

#P17655

Entrez-Gene Id:

824

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:50 - 1:100
Immunofluorescence (Immunocytochemistry) 1:200 - 1:400

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

Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb recognizes endogenous levels of total calpain 2 (large subunit) protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human calpain 2 (large subunit) protein.

Background

Calpain is a calcium-dependent thiol proteinase that is functionally active as a heterodimer composed of a small regulatory subunit and one of at least two large catalytic subunits (calpain 1 or calpain 2). In vitro, calpain 1 (mu-calpain) requires micromolar levels of calcium, while calpain 2 (M-calpain) requires millimolar levels of calcium for activation. The regulation of calpain in vivo is the subject of many current studies, which suggest that proteolytic activity is regulated post-transcriptionally by mechanisms such as calcium requirements, subcellular localization of the heterodimer, phosphorylation via the EGFR-Erk signaling cascade, endogenous inhibitors (calpastatin), and autoproteolytic cleavage (1). Calpastatin negatively regulates autoproteolytic cleavage of calpain 1 between Gly27 and Leu28 (2). Calpain influences cell migration by modifying rather than degrading its substrates responsible for cell adhesion and cytoskeletal arrangement. Control of calpain activity has caught the attention of drug development since limiting its activity could mute invasiveness of tumors or chronic inflammation (1).

  1. Perrin, B.J. and Huttenlocher, A. (2002) Int. J. Biochem. Cell Biol. 34, 722-725.
  2. Melloni, E. et al. (1996) Biochem. Biophys. Res. Commun. 229, 193-197.

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 nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) 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.
XP is a registered 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

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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
#70655

Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb

Western Blotting Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb(上图)或 β-Actin (D6A8) Rabbit mAb #8457(下图)对各种细胞系的提取物进行蛋白质印迹分析。
Immunoprecipitation Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
对来自 PC-3 细胞提取物的人钙蛋白酶 2(大亚基)蛋白进行免疫沉淀分析。泳道 1 为 10% input,泳道2为 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900,泳道 3 为 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb。使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb 进行蛋白质印迹分析。Anti-rabbit IgG, HRP-linked Antibody #7074 用作二抗。
Immunofluorescence Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
对使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb(红色)标记以及使用 DAPI #4083(蓝色)共同标记的固定冷冻的小鼠海马进行共聚焦免疫荧光分析。
Immunofluorescence Image 2: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
对使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb(红色)标记以及使用 DAPI #4083(蓝色)共同标记的固定冷冻的小鼠结肠进行共聚焦免疫荧光分析。
Immunofluorescence Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb(绿色)、β-Actin (8H10D10) Mouse mAb #3700(红色)和 DAPI #4083(蓝色)对模拟转染(左图,高表达)或用针对人钙蛋白酶 2(大亚基)(右图,低表达)的 siRNA 转染的 C2C12 细胞进行共聚焦免疫荧光分析。
Immunofluorescence Image 2: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
使用 Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb(绿色)、β-Actin (8H10D10) Mouse mAb #3700(红色)和 DAPI #4083(蓝色)对模拟转染(左图,高表达)或用针对人钙蛋白酶 2(大亚基)(右图,低表达)的 siRNA 转染的 PC-3 细胞进行共聚焦免疫荧光分析。