The antiserum against calretinin is produced in rabbits by immunization with recombinant human calretinin containing a 6-his tag at the N-terminal. The antibody was evaluated for specificity and potency: a) by Biotin-Avidin labeling of cryostate-, vibratome- and paraffinsections of 4% paraformaldehyde fixed brains and b) by immunoenzymatic labelling of immunoblots。
稀釋
Immunohistochemistry: 1:2'000 - 1:5'000, on paraformaldehyde (4%) or formalin-fixed tissue, cryostate or paraffin-sections.
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. Schwaller B., Buchwald P., Blümcke I., Celio M.R. and Hunziker W. (1994) Characterization of a polyclonal antiserum against the purified human recombinant calcium-binding protein calretinin. Cell Calcium 14: 639-648.
2. Schiffmann S.N. et al (1999) Impaired motor coordination and Purkinje cell excitability in mice lacking calretinin. PNAS, 96: 5257-5262.
3. Gotzos V., Vogt P. and M.R. Celio (1995) Calretinin is a selective marker for malignant pleural mesotheliomas of the epithelial type. Pathol. Res. Pract. 192:137- 147.
4. Doglioni, C. et al. (1996) Calretinin: a novel immunocytochemical marker for mesothelioma. Am. J. Surg. Pathol. 20:1037-1046.