This antibody does not cross react with the other calpain family members (calpain 1, calpain 2, calpain 3, etc.) It recognizes latent and amino-processed Calpain 10.
This antibody binds to the middle of the catalytic domain, aminoterminal from the catalytic histidine.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Detects a band of approximately 78 66 kDa (predicted molecular weight: 75 kDa). A recommended starting concentration for Western blots is 1/1000 when using colorimetric substrates such as BCIP/NBT, and 1/5000 for chemiluminescent substrates. Dilution optimised using Chromogenic detection.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Calcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
Involvement in disease
Defects in CAPN10 are a cause of susceptibility to diabetes mellitus non-insulin-dependent type 1 (NIDDM1) [MIM:601283]. It is a multifactorial disorder of glucose homeostasis caused by a lack of sensitivity to the body's own insulin. Affected individuals usually have an obese body habitus and manifestations of a metabolic syndrome characterized by diabetes, insulin resistance, hypertension and hypertriglyceridemia. The disease results in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
Belongs to the peptidase C2 family. Contains 1 calpain catalytic domain.
IHC image of ab28220 staining in human normal pancreas formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab28220, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.