Immunofluorescent analysis of Calpain in HeLa cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a Calpain monoclonal antibody (Product # MA3-943) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). Calpain staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Images were taken at 60X magnification.
|Tested species reactivity||Bovine, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified bovine skeletal muscle 28 kDa calpain subunit.|
|Storage buffer||ascites diluted in PBS|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunofluorescence (IF)||1:10 - 1:100|
|Western Blot (WB)||1:1000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA3-943 detects calpain-28 kDa subunit from bovine platelet, skeletal and heart muscle and human tissues. This antibody does not cross-react with the human 80 kDa m-calpain subunit.
MA3-943 has been successfully used in Western blot and immunofluorescence procedures. By Western blot, this antibody detects a 28 kDa protein representing the 28 kDa subunit of calpain from bovine heart extract.
The MA3-943 antigen is purified bovine skeletal muscle 28 kDa calpain subunit.
The calpain (calcium-dependent protease or calcium-activated neutral protease) system consists of two ubiquitous forms of calpain (mu-calpain and m-calpain), a tissue specific calpain (n-calpain), and a calpain inhibitory protein (calpastatin). The calpain system has been detected in every vertebrate tissue examined, and has been suggested to play a regulatory role in cellular protein metabolism. This regulatory role may have important implications in platelet aggregation and pathologies associated with altered calcium homeostasis and protein metabolism such as ischemic cell injury and degenerative diseases. Inhibitors of calpain have been shown to block dexamethasone and low-level irradiation induced apoptosis in thymocytes suggesting that calpain has a regulatory or mechanistic role in apoptotic cell death.
Mu- and m-calpains are heterodimers consisting of 28 kDa and 80 kDa subunits. The 28 kDa subunit is identical in the two isoforms, but the 80 kDa subunits differ with ~50% sequence similarity. 28 kDa/80 kDa complexes are thought to be inactive proenzymes which, upon binding of calcium, undergo conformational changes that promotes cleavage of the 28 kDa subunit and results in enzyme activation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.