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Merck
CN

MAB3084-C

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

clone PI-11, from mouse

别名:

Calpastatin, Calpain inhibitor, Sperm BS-17 component

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关于此项目

eCl@ss:
32160702
UNSPSC Code:
12352203
Clone:
PI-11, monoclonal
Species reactivity:
human
Application:
ICC, IF, IHC, WB
Citations:
9
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biological source

mouse

antibody form

purified immunoglobulin

antibody product type

primary antibodies

clone

PI-11, monoclonal

species reactivity

human

species reactivity (predicted by homology)

rat (based on 100% sequence homology)

technique(s)

immunocytochemistry: suitable, immunofluorescence: suitable, immunohistochemistry: suitable (paraffin), western blot: suitable

isotype

IgG1κ

NCBI accession no.

UniProt accession no.

shipped in

dry ice

target post-translational modification

unmodified

Gene Information

human ... CAST(831)

General description

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.

Immunogen

Human erythrocyte calpastatin.

Application

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.

Biochem/physiol Actions

Reacts with human calpastatin.

Physical form

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

Preparation Note

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.

Analysis Note

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.

Other Notes

Concentration: Please refer to lot specific datasheet.
Replaces: MAB3084

Disclaimer

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.

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存储类别

12 - Non Combustible Liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

法规信息

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Masaya Takahashi et al.
Journal of cardiovascular pharmacology, 47(3), 413-421 (2006-04-25)
Changes in proteolytic activity of the myocardium during the development of heart failure after left coronary artery ligation (CAL) of rats were examined. Hemodynamics of the rats at the eighth week (8w-CAL rat), but not at the second week (2w-CAL
Masaya Takahashi et al.
Cardiovascular research, 65(2), 356-365 (2005-01-11)
Genetic depletion of dystrophin-related protein (DRP) complex causes cardiomyopathy in animals and humans. We found in a previous study that some types of DRP were degraded and that calpain content was increased in rats with non-genetically induced heart failure. The
Robert W Neumar et al.
The Journal of biological chemistry, 278(16), 14162-14167 (2003-02-11)
Cross-talk between calpain and caspase proteolytic systems has complicated efforts to determine their distinct roles in apoptotic cell death. This study examined the effect of overexpressing calpastatin, the specific endogenous calpain inhibitor, on the activity of the two proteolytic systems
Hiroyuki Yoshida et al.
Cardiovascular research, 59(2), 419-427 (2003-08-12)
Genetic defects in several sarcoglycans (SGs) and dystrophin (Dys) play a critical role in cardiomyopathy. The present study was designed to determine whether changes in SGs and Dys might occur in animals with chronic heart failure (CHF) induced by acute
Yanzhang Li et al.
Experimental neurology, 220(2), 316-319 (2009-09-16)
Calpains and caspases are cysteine endopeptidases which share many similar substrates. Caspases are essential for caspase-dependent apoptotic death where calpains may play an augmentive role, while calpains are strongly implicated in necrotic cell death morphologies. Previous studies have demonstrated a

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