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CaF2-5D2 INVESTIGATOR Name Douglas M. Fambrough Address Department of Biology, The Johns Hopkins University, 34th & Charles Streets, Baltimore, MD 21218 E-Mail [email protected] IMMUNOGEN Substance Name Origin Chemical Composition Developmental Stage IMMUNIZATION PROTOCOL Donor Animal Species Strain Sex Organ and tissue Immunization Dates immunized Amount of antigen Route of immunization Adjuvant FUSION Date Myeloma cell line Species Designation MONOCLONAL ANTIBODY Isotype Specificity Cell binding Immunohistology Antibody competition Species Specificity chicken pectoralis muscle SR vesicle prep chicken SR membranes adult mouse BALB/cJ spleen 200 g SR prep intraperitoneal; tail vein boost 7, 14, & 21 days later Freund's complete mouse SP2/0 Ag8 IgG1, kappa light chain B tryptic fragment (45 kDa) of SERCA1 (fast twitch SR Ca+2-ATPase) all vertebrates tested to date ANTIGEN Chemical properties Ca+2 SR ATPase fast twitch muscle Molecular weight 110 kDa Characterization Immunoprecipitation Immunoblotting 0.5 g/ml very good Purification Amino acid sequence analysis Functional effects unknown Immunohistochemistry very good on cryosections and formaldehyde or methanol fixed tissues 5 g/ml PUBLICATIONS : Airey, J., Baring, M., Beck, C., Chelliah, Y., Deerinck, T., Ellisman, M., Houenou, L., McKemy, D., Sutko, J., and Talvenheimo, J.(1993). Failure to make normal ryanodine receptor is an early event associated with the crooked neck dwarf (cn) mutation in chicken. Devel. Dyn. 197, 169-188. Airey, J., Deerinck, T., Ellisman, M., Houenou, L., Ivaneko, A., Kenyon, J., McKemy, D., and Sutko, J. (1993). Crooked neck dwarf (cn) mutant chicken skeletal muscle cells in low density primary cultures fail to express normal alpha ryanodine receptor and exhibit a partial mutant phenotype. Devel. Dyn. 197, 189-202. (Continued) CaF2-5D2 (Continued) Murphy, R.M., Larkins, N.T., Mollica, J.P., Beard, N.A., and Lamb, G.D. (2009). Calsequestrin content and SERCA determine normal and maximal Ca2+ storage levels in sarcoplasmic reticulum of fast- and slow-twitch fibres of rat. J. Physiol. 587.2, 443-460. Murphy, R.M., and Lamb, G.D. (2009). Endogenous Calpain-3 activation is primarily governed by small increases in resting cytoplasmic [Ca2+] and is not dependent on stretch. J. Biol. Chem. 284(12), 7811-7819. ACKNOWLEDGMENTS STATEMENT We have been asked by NICHD to ensure that all investigators include an acknowledgment in publications that benefit from the use of the DSHB's products. We suggest that the following statement be used: “The (select: hybridoma, monoclonal antibody, or protein capture reagent,) developed by [Investigator(s) or Institution] was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at The University of Iowa, Department of Biology, Iowa City, IA 52242.” Please send copies of all publications resulting from the use of Bank products to: Developmental Studies Hybridoma Bank Department of Biology The University of Iowa 028 Biology Building East Iowa City, IA 52242