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.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

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
Western blot analysis of extracts from various cell lines using Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Immunoprecipitation Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
Immunoprecipitation of human calpain 2 (large subunit) protein from PC-3 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb. Western blot analysis was performed using Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.
Immunofluorescence Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse hippocampus, labeled with Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb (red) and co-labeled with DAPI #4083 (blue).
Immunofluorescence Image 2: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse colon, labeled with Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb (red) and co-labeled with DAPI #4083 (blue).
Immunofluorescence Image 1: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of C2C12 cells, mock transfected (left, high-expressing) or transfected with siRNA directed against human calpain 2 (large subunit) (right, low-expressing), using Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
Immunofluorescence Image 2: Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of PC-3 cells, mock transfected (left, high-expressing) or transfected with siRNA directed against human calpain 2 (large subunit) (right, low-expressing), using Calpain 2 Large Subunit (M-type) (E3M6E) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).