Certificate of Analysis and Data Sheet

Source:
Human Eryhtrocytes

Catalog No.
PRO-363

Description:

This protein consists of an 80-kDa large subunit and a 30 kDa small subunit.
The protein was purified by sequential chromatography through DEAE-Sepharose, A1.5m Bio-Gel, and Phenyl-Sepharose CL-4B columns.

Physical Appearance:

Sterile Filtered colorless solution.

Formulation:

50mM imidazole-HCl, 100mM NaCl, 5mM EGTA, 1mM DTT and 10% sucrose.

Stability:

?Calpain although stable at 10⁰C for 1 week, should be stored desiccated below -18⁰C.
For long term storage it is recommended to add a carrier protein (0.1% HSA or BSA).
Please avoid freeze-thaw cycles.

Purity:

Greater than 90% as determined by:
Analysis by reducing and non-reducing SDS-PAGE Coomassie.

Applications:

This protein can be used for immunoblots, absorption experiments in immunohistochemistry, radioimmunoassay and intracellular injection.
For adsorption we suggest the following procedure:
A- Dilute 1痞 of the antiserum against ?calpain in 1ml of the usual buffer for immunohistochemistry (final dilution 1:1000).
B- Add 1ug of protein to 1ml of the diluted antibody solution and mix well.
C- Incubate for at least 6 hours in the cold.
D- Apply to tissue-sections and incubate for 3 days.
E - Complete the immunohistochemical reaction as usual (biotinylated second antibody, ABC-complex, DAB).
As a result, the immunostaining should be strongly reduced or even completely prevented.

Refrencesy:

Lane et al. (1992) A comparison of the intracellular distribution of ?calpain, mcalpain and calpastatin in proliferating human A431 cells. Exp. Cell Res. 203:5-16.

Usage:

This material is offered by ProSpec-TechnoGene for research, laboratory or further evaluation purposes.

Gene:

Name:CAPN1
Synonyms:CANPL1

Protein synonyms/aliases:

Calpain-1 catalytic subunit (EC 3.4.22.52)
    (Calpain-1 large subunit)
    (Calcium-activated neutral proteinase 1)
    (CANP 1)
    (Calpain mu-type)
    (muCANP)
    (Micromolar-calpain).

Protein Family:

Belongs to the peptidase C2 family.

Protein Domains:

Domains:
•IPR001300   Peptidase C2, calpain
•IPR002048   Calcium-binding EF-hand

Protein links:

Precursor- Protein structure and amino acid sequence:

Latest Publications:

1. Increased expression of glial fibrillary acidic protein fragments and mu-calpain activation within the hippocampus of prion-infected mice.
Biochem Soc Trans 2006 Feb;34(Pt 1):51-4
2. Mitochondrial localization of mu-calpain.
Biochem Biophys Res Commun 2005 Dec 16;338(2):1241-7
3. Determination of peptide substrate specificity for mu-calpain by a peptide library-based approach: the importance of primed side interactions.
J Biol Chem 2005 Dec 9;280(49):40632-41
4. mu-Calpain and calpain-3 are not autolyzed with exhaustive exercise in humans.
Am J Physiol Cell Physiol 2006 Jan;290(1):C116-22
5. mu-Calpain regulates receptor activator of NF-kappaB ligand (RANKL)-supported osteoclastogenesis via NF-kappaB activation in RAW 264.7 cells.
J Biol Chem 2005 Aug 19;280(33):29929-36
6. Mu-calpain is functionally required for alpha-processing of Alzheimer's beta-amyloid precursor protein.
Biochem Biophys Res Commun 2005 May 13;330(3):714-21

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中華民國95年06月06日更新