Anti-Calpastatin Antibody, clone PI-11 (Ascites Free)

Calpastatin (UniProt P20810; also known as Calpain inhibitor, Sperm BS-17 component) is encoded by the CAST gene (Gene ID 831) in human. Calpastatin is a ubiquitously expressed protein that functions as the endogenous inhibitor of both calpain 1 and 2 (micro-calpain and m-calpain, respectively). Calpastatin contains four inhibitory repeats, each of which neutralizes an activated calpain with exquisite specificity and potency. Fully activated calpain binds ten Ca(2+) ions, resulting in several conformational changes allowing recognition by calpastatin. Calpain inhibition is mediated by the intimate contact with three critical regions of calpastatin. Two regions target the penta-EF-hand domains of calpain and the third occupies the substrate-binding cleft, projecting a loop around the active site thiol to evade proteolysis. The inhibitory domain of calpastatin recognizes multiple lower affinity sites present only in the calcium-bound form of the enzyme, resulting in an interaction that is tight, specific and calcium dependent.

~110/120 kDa observed. Target band(s) appear larger than the calculated molecular weights due to posttranslational modifications.

Reacts with human calpastatin.

Human erythrocyte calpastatin.

Immunohistochemistry Analysis: A 1:250 dilution from a representative lot detected Calpastatin in human prostate cancer tissue, as well as normal testis and kidney tissues.
Immunohistochemistry Analysis: A representative lot detected both cytoplasmic and nuclear calpastatin immunoreactivity in cancer cells using paraffin-embedded pancreatic adenocarcinoma, as well as bile duct and ampullary tumour tissue sections (Storr, S.J., et al. (2012). BMC Cancer. 12:511).
Immunohistochemistry Analysis: A representative lot detected the exogenously expressed human calpastatin, but not the endogenous rat calpastatin, in frozen rat heart tissue sections (Maekawa, A., et al. (2003). J. Mol. Cell. Cardiol. 35(10):1277-1284).
Immunocytochemistry Analysis: A representative lot detected exogenously expressed human calpastatin, but not the endogenous rat calpastatin, in primary rat cortical neurons by fluorescent immunocytochemistry (Ma, M., et al. (2012). J. Neurotrauma. 29(16): 2555–2563).
Immunocytochemistry Analysis: A representative lot detected calpastatin intracellular localization in proliferating A431 cells (Lane, R.D., et al. (1992), Exp. Cell. Res. 203(1):5-16).
Immunofluorescence Analysis: A representative lot detected the exogenously expressed human calpastatin, but not the endogenous rat calpastatin, in retinal ganglion cells (RGCs) by fluorescent immunohistochemical staining of retinal flat mounts (Ma, M., et al. (2012). J. Neurotrauma. 29(16): 2555–2563).
Western Blotting Analysis: Representative lots detected exogenously expressed human calpastatin, but not the endogenous rat calpastatin, in rat optical nerve and heart tissue homogenates (Ma, M., et al. (2012). J. Neurotrauma. 29(16): 2555–2563; Maekawa, A., et al. (2003). J. Mol. Cell. Cardiol. 35(10):1277-1284).
Western Blotting Analysis: A representative lot detected a 75% increase of rat brain calpastatin level during development from E18 to P90 (Li. Y., et al. (2009). Exp. Neurol. 220(2):316-319).
Western Blotting Analysis: Representative lots detected endogenous calpastatin in rat heart tissue homogenates (Takahashi, M., et al. (2006). J. Cardiovasc. Pharmacol. 47(3):413-421; Takahashi, M., et al. (2005). Cardiovasc. Res. 65(2):356-365; Yoshida, H., et al. (2003). Cardiovasc. Res. 59(2):419-427).
Western Blotting Analysis: A representative lot detected calpastatin in lysates from untransfected SH-SY5Y cells as well as transfected SH-SY5Y cells overexpressing human calpastatin (Neumar, R.W., et al. (2003). J. Biol. Chem. 278(16):14162-14167).

Research Category
Neuroscience

Research Sub Category
Developmental Neuroscience

This Anti-Calpastatin Antibody, clone PI-11 (Ascites Free) is validated for use in Western Blotting, Immunohistochemistry (Paraffin), Immunocytochemistry and Immunofluorescence for the detection of Calpastatin.

Replaces: MAB3084

Protein G Purified

Format: Purified

Purified mouse monoclonal IgG1κ antibody in buffer containing 0.1 M Tris-Glycine (pH 7.4), 150 mM NaCl without preservatives.

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Evaluated by Western Blotting in MDA-MB-231 cell lysate.

Western Blotting Analysis: 0.5 µg/mL of this antibody detected Calpastatin in 10 µg of MDA-MB-231 cell lysate.

Concentration: Please refer to lot specific datasheet.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.