Application

Research CategorySignaling

Research Sub CategorySignaling 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).

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.

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.

General description

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.

Immunogen

Purified m-calpain from rabbit skeletal muscle.

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

Mouse monoclonal IgG2aκ ascites with 0.05% sodium azide.

Unpurified

Quality

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.

Specificity

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.

Storage and Stability

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.

Target description

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