Immunofluorescent analysis of M-Calpain in MCF-7 Cells. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with a M-Calpain monoclonal antibody (Product # MA3-942) at a dilution of 1:20 overnight at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503). M-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, Pig, Rabbit, Rat|
|Published species reactivity||Yeast, Rabbit, Bovine, Fish, Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified bovine skeletal muscle 80 kDa subunit of m-calpain.|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay dependent|
|Flow Cytometry (Flow)||Assay Dependent|
|Immunohistochemistry (Frozen) (IHC (F))||1:50|
|Western Blot (WB)||1:500|
* 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.
MA3-942 detects the 80 kDa subunit of m-calpain in human platelets and erythrocytes, bovine platelets, heart and skeletal muscle, rat myoblasts, kidney, liver and spleen, pig cultured cells, and rabbit samples. This antibody does not cross-react with mu-calpain, n-calpain, calmodulin or calpastatin.
MA3-942 has been successfully used in Western blot, IF, immunohistochemistry, and ELISA procedures. By Western blot, this antibody detects an ~80 kDa protein representing m-calpain in bovine platelets. Immunocytochemical staining of m-calpain in LLC-PK1 cells with MA3-942 results in diffuse cytoplasmic staining. This product has not been shown to be effective in immunoprecipitation experiments.
The MA3-942 antigen is purified bovine skeletal muscle 80 kDa subunit of m-calpain. This antibody recognizes an epitope between amino acids 502-699 (domain III/IV) of human calpain.
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-Calpain, also known as Calpain-I, and m-calpain, also known as Calpain-II, are intracellular, calcium-dependent cysteine proteases.
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.
Role of mu-calpain in human decidua for recurrent miscarriage.
MA3-942 was used in western blot to study the role of mu-calpain in decidua from patients with recurrent miscarriage
|Kumagai K,Ozaki Y,Nakanishi T,Inomata M,Furuno T,Nakanishi M,Ogasawara MS||American journal of reproductive immunology (New York, N.Y. : 1989) (59:339)||2008|
Purification and characterization of calpain and calpastatin from rainbow trout, Oncorhynchus mykiss.
MA3-942 was used in western blot to purify and characterize the calpain and calpastatin from rainbow trout
|Saito M,Li H,Thompson VF,Kunisaki N,Goll DE||Comparative biochemistry and physiology. Part B, Biochemistry and molecular biology (146:445)||2007|
Effects of 25-hydroxyvitamin D3 and manipulated dietary cation-anion difference on the tenderness of beef from cull native Korean cows.
MA3-942 was used in western blot to study the effect of 25-hydroxyvitamin D3 and manipulated dietary cation-anion difference on beef quality
|Cho YM,Choi H,Hwang IH,Kim YK,Myung KH||Journal of animal science (84:1481)||2006|
Calpain may contribute to hereditary cataract formation in sheep.
MA3-942 was used in western blot to investigate the role of calpain during hereditary cataract formation in sheep
|Robertson LJ,Morton JD,Yamaguchi M,Bickerstaffe R,Shearer TR,Azuma M||Investigative ophthalmology and visual science (46:4634)||2005|
Changes in the calpains and calpastatin during postmortem storage of bovine muscle.
MA3-942 was used in western blot to detect the changes of micro-calpain, m-calpain, and calpastatin in bovine muscle during postmortem storage
|Boehm ML,Kendall TL,Thompson VF,Goll DE||Journal of animal science (76:2415)||1998|
Effect of monoclonal antibodies specific for the 28-kDa subunit on catalytic properties of the calpains.
MA3-942 was used in western blot to study the effect of specific calpain antibodies for the regulation of calpain function
|Cong J,Thompson VF,Goll DE||The Journal of biological chemistry (268:25740)||1993|
Fiber type-specific expression of major proteolytic systems in fast- to slow-transforming rabbit muscle.
MA3-942 was used in immunohistochemistry to study the importance of two major proteolytic systems in tranforming rabbit and rat muscles
|Sultan KR,Dittrich BT,Leisner E,Paul N,Pette D||American journal of physiology. Cell physiology (280:C239)||2001|