Specificityab28257 recognizes the amino terminal end of the large subunit of Calpain 1. The antibody does not recognize amino-processed Calpain 1, and can be used to discriminate between latent and amino-processed forms. Also, it does not cross react with the other calpain family members (M Calpain, Calpain 94, ncl2, ncl3, etc.).
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 82 kDa (predicted molecular weight: 82 kDa). 1/1000, when using colorimetric substrates such as BCIP/NBT - 1/5000, when using chemiluminescent substrates. But note that when used against the reduced protein, ab28257 identifies bands at 80 kDa, 68 kDa, and a series of further cleaved forms. Dilution optimised using Chromogenic detection.
Use a concentration of 1 - 5 µg/ml.
FunctionCalcium-regulated non-lysosomal thiol-protease which catalyze limited proteolysis of substrates involved in cytoskeletal remodeling and signal transduction.
Sequence similaritiesBelongs to the peptidase C2 family. Contains 1 calpain catalytic domain. Contains 4 EF-hand domains.
Cellular localizationCytoplasm. Cell membrane. Translocates to the plasma membrane upon Ca(2+) binding.
Calcium activated neutral proteinase small subunit antibody
Calcium dependent protease small subunit 1 antibody
Calcium dependent protease small subunit antibody
Calcium-activated neutral proteinase 1 antibody
Calpain 1 antibody
Calpain 1 large subunit antibody
Calpain 1, (mu/I) large subunit antibody
Calpain mu type antibody
Calpain mu-type antibody
Calpain regulatory subunit antibody
Calpain small subunit 1 antibody
Calpain, large polypeptide L1 antibody
Calpain-1 catalytic subunit antibody
Calpain-1 large subunit antibody
CANP 1 antibody
CANP small subunit antibody
CANPL 1 antibody
CAPN 1 antibody
Cell proliferation inducing protein 30 antibody
Cell proliferation-inducing gene 30 protein antibody
Micromolar Calpain antibody
Mu Calpain antibody
Anti-Calpain 1 antibody images
Immunocytochemistry/ Immunofluorescence-Calpain 1 antibody - Aminoterminal end domain I(ab28257)
ICC/IF image of ab28257 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab28257, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot - Calpain 1 antibody - Aminoterminal end domain I (ab28257)
All lanes : Anti-Calpain 1 antibody (ab28257) at 1 µg/ml
Lane 1 : Human liver tissue lysate - total protein (ab29889) Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 82 kDa Observed band size : 82 kDa Additional bands at : 27 kDa,42 kDa,65 kDa. We are unsure as to the identity of these extra bands.