Anti-Calpain I/II, large subunit Antibody, clone 1D10A7

Calpains I & II are intracellular cysteine proteinases distributed in various tissues and cells. Calpain I & II are also known as mu- and m-calpains, respectively, based on their calcium requirement, with calpain I being active at micromolar Ca+2 concentrations and calpain II at millimolar Ca+2 concentrations. Both calpains are composed of a large catalytic (80 kDa) and a small regulatory (30 kDa) subunit, and each subunit contains a calmodulinlike domain at the C-terminus. Upon Ca2+ binding to the calmodulin-like domains, calpains become activated for catalyzing both substrate proteolytic degradation as well as autolysis. Fourteen calpain large subunits have been identified in human (calpain 1-3 & 5-15 encoded by CAPN1-3 & 5-15 gene) and two small subunits encoded by the CAPNS1 & CAPSN2 genes. The calpain small subunit 1 encoded by CAPNS1 is also known as calpain 4. To date, calpain I & II remain the best characterized members of the calpain family. Calpain activity was initially characterized as “calcium-activated neutral protease” (CANP) detected in brain, lens of the eye, and other tissues. The name “calpain” was later adopted as a hybrid name from two well-known proteins at the time, calmodulin and papain.

~80 kDa observed. Uncharacterized band(s) may appear in some lysates.

Purified m-calpain from rabbit skeletal muscle.

Detect CAPN1 using this mouse monoclonal Anti-Calpain I/II, large subunit Antibody, clone 1D10A7, Cat. No. MABS1215, validated for use in Western Blotting and Immunocytochemistry.

Research Category
Signaling

Research Sub Category
Signaling Neuroscience

Western Blotting Analysis: A 1:20,000 dilution from a representative lot detected recombinant human calpain-2 and calpain-1 C115S mutant protein.
Western Blotting Analysis: A 1:20,000 dilution from a representative lot detected Calpain, large subunit in rat spleen tissue lysate.
Immunocytochemistry Analysis: A representative lot detected Calpain, large subunit in mouse embryonic fibroblast (MEF) cells (Yamada, M., et al. (2009). Nat Med. 15(10):1202-1207).
Western Blotting Analysis: A representative lot detected Calpain, large subunit in cytosolic preparation from rat cortical tissue (Blomgren, K., et al. (1995). Brain Res. 684(2):143-149).
Western Blotting Analysis: A representative lot detected purified mu- and m-calpain large subunits from rabbit, monkey, human, and rat tissues (Kawashima, S., et al. (1998). Biol Chem. 379(2):201-204).
Western Blotting Analysis: A representative lot detected Calpain, large subunit in normal & cataractous rat lens homogenates (Inomata, M., et al. (2009). Nat Med. 15(10):1202-1207).
Western Blotting Analysis: A representative lot detected Calpain, large subunit in MEF cells (Yamada, M., et al. (2009). Nat Med. 15(10):1202-1207).

Clone 1D10A7 detects the large catalytic (80K), but not the small regulatory (30K), subunit of mu-calpain (calpain I) and m-calpain (calpain II), with ~7-fold higher affinity toward m-calpain than mu-calpain.

Unpurified

Mouse monoclonal IgG2aκ ascites with 0.05% sodium azide.

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 rat lung tissue lysate.

Western Blotting Analysis: A 1:20,000 dilution of this antibody detected Calpain, large subunit in rat lung tissue lysate.

Concentration: Please refer to lot specific datasheet.

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