Product Pathways - Protein Stability
Calpain 2 Large Subunit (M-type) Antibody #2539
PhosphoSitePlus® protein, site, and accession data: calpain 2
| Applications | Reactivity | Sensitivity | MW (kDa) | Source |
|---|---|---|---|---|
| W IP IF-F F | H M R | Endogenous | 80 | Rabbit |
Applications Key:
W=Western Blotting
IP=Immunoprecipitation
IF-F=Immunofluorescence (Frozen)
F=Flow Cytometry
Reactivity Key:
H=Human
M=Mouse
R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
Specificity / Sensitivity
Calpain 2 Large Subunit (M-type) Antibody detects endogenous levels of total calpain 2 (large subunit) protein. The antibody detects full-length calpain 2 as well as calpain 2 autoproteolytically cleaved at serine 20. The antibody does not detect recombinant calpain 1.
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the human sequence of calpain 2 (large subunit). Antibodies are purified by protein A and peptide affinity chromatography.
Western Blotting
Western blot analysis of extracts from C2C12 and PC3 cells, using Calpain 2 Large Subunit (M-type) Antibody.
Flow Cytometry
Flow cytometric analysis of C2C12 cells, using Calpain 2 Large Subunit (M-type) Antibody (blue) compared to a nonspecific negative control antibody (red).
IF-F
Confocal immunofluorescent analysis of mouse retina using Calpain 2 Large Subunit (M-type) Antibody (red) and Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411 (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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).
- Perrin, B.J. and Huttenlocher, A. (2002) Int. J. Biochem. Cell Biol. 34, 722-725.
- Melloni, E. et al. (1996) Biochem. Biophys. Res. Commun. 229, 193-197.
Application References
- Beltran, L. et al. (2011) Proc Natl Acad Sci U S A 108, 16217-22. Applications: IP Western Blotting
- Pilop, C. et al. (2009) Circulation 120, 983-91. Applications: Western Blotting
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Companion Products
- 2556 Calpain 1 Large Subunit (Mu-type) Antibody
- 2232 EGF Receptor Antibody
- 3285 FAK Antibody
- 9911 Phospho-Erk1/2 Pathway Sampler Kit
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
For Research Use Only. Not For Use In Diagnostic Procedures.
