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Rabbit Anti-Calpain-12
Amino Terminal End Domain-IV, large subunit
CL4CALPAIN12
Lot:
Description:
Cedarlane’s polyclonal antibody to the cysteine protease Calpain-12 is made in rabbits, using a synthetic peptide based
on domain-IV in the large subunit using the mouse sequence. The antibody binds to Calpain-12, but does not cross react
with the other calpain family members (Calpain-1, Calpain-2, ncl-2, ncl-3 etc.).
Formulation:
This antibody is provided in a 100μg size. It has been peptide-affinity purified and concentrated to 1 mg/ml, with the
addition of 0.05% Sodium Azide as preservative, and 50% glycerol as cryoprotectant.
Stability:
The undiluted antibody solution is stable for short term at 4°C. For longer storage, store at -20°C. Avoid unnecessary
freeze-thaw cycles. Diluted antibody can be reused several times if not contaminated.
Applications:
Calpain-12 is an intracellular, calcium-dependent cysteine protease. Calpain-12 was first described in the mouse (most
strongly in the skin), and maps to mouse chromosome 7. The 720 amino acid sequence predicts a protein of 80.588 kD,
and the truncated 502, 462 and 449 amino acid sequences predict proteins of 55.874, 51.492 and 50.397 kD respectively.
The different isoforms differ in the carboxyterminal ends, ending with aberrant domain-III and lacking domain IV.
Domains in the large subunit include the amino terminal domain-I, the proteinase domain-II, domain-III, and the EF-hand
domain-IV, making Calpain-12 most similar to the "classical" calpains 1 & 2. Sequence alignments place Calpain-12
closest to chicken calpain u/m, and it may be that Calpain-12 is the human orthologue of this protein.
This antibody recognizes latent and the aminotruncated (active) Calpain-12, and recognizes only the longest (720 AA)
form of calpain-12. The latent large subunit migrates at about 80 kD on reduced SDS PAGE gels, and truncation at the
amino terminal end yields an approximately 58 kD form. A cascade of smaller forms can be seen with further activation.
CL4CALPAIN12 works well for Western Blotting applications, and when used against the reduced protein identifies
bands at 80 kD, and a series of further cleaved forms. It will also bind to the non-reduced protein, and thus may be
suitable for immunoprecipitation, immunohistochemistry, and ELISA. Not all cell types produce Calpain-12; since it is a
cellular protein, cell lysate works well for Western Blots. Rat, mouse and human Calpain-12 are detected with this
antibody. A recommended starting concentration for Western Blots is 1:1000 when using colorimetric substrates such as
BCIP/NBT, and 1:5000 for chemiluminescent substrates. Higher concentrations of antibody may be needed for samples
from more distantly related species.
FOR RESEARCH USE ONLY.
JJ 01/11/07