The antiserum against Ca2+-ATPase, isoform 2, was produced in rabbits by immunization with the N-terminal sequence-specific peptide of the human pump as epitope. It recognizes the corresponding proteins in human, rat and mouse tissue. The antibody was evaluated for specificity and potency : a) by Biotin-Avidin labelling of cryostatesections of 4% paraformaldehyde fixed tissue and b) by immunoenzymatic labelling of Western blots of brain membranes or primary nerve cell cultures. It can also be used for immunoprecipitation. The serum was inactivated at 50°C for 30 minutes.
稀釋
Immunohistochemistry : 1: 1'000 - 1: 2'000.
Immunoblots : 1: 1'000 - 1: 2'000.
For immunohistochemistry and immunoblots the titer was determined by using the avidin-biotin method. We recommend that the optimal dilutions be determined by titration experiments.
儲存
After reconstitution freeze in small aliquots (e.g. 1 μl) and keep at - 80°C (or at least - 20°C). For continuous use keep at 4°C (with 0.01% Na-azide). Avoid repeated freezing and thawing.
參考文獻
1. Stauffer T.P., Guerini D., Carafoli E. (1995) Tissue distribution of the four gene products of the plasma-membrane Ca2+-pump. A study using specific antibodies. J. Biol. Chem. 19: 12184- 12190. 2. Stauffer T.P., Guerini D., Celio M.R., Carafoli E. (1997) Immunolocalization of the plasma membrane Ca2+-pump isoforms in the rat brain. Brain Res. 748:21-29.