L7/L12 | Ribosomal protein
AS08 331 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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Purified native Chlamydomonas reinhardtii L-30 protein eluted from a gel piece
11.9 kDa (Chlamydomonas reinhardtii), 15.7 kDa (spinach)
Total protein extracted freshly from Chlamydomonas reinhardtii denatured ay 90°C 5 min. were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blot was blocked with % milk or % BSA for 1h/RT or 4°C/ON with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed chemiluminescent detection reagent, according to manufacture's recommendations.
Name of this antibody has been changed from L-30 | 50S ribosomal protein L30 to L7/L12 based on the following reference: Randolph-Anderson et al. (1989). Electrophoretic and immunological comparisons of chloroplast and prokaryotic ribosomal proteins reveal that certain families of large subunit proteins are evolutionarily conserved. J Mol Evol. 1989 Jul;29(1):68-88.
Cross react with L2 and L26 proteins of Chlamydomonas reinhardtii. L7/L12 is very acidic, may not bind well to nitrocellulose membrane and can have abberant mobility depending upon conditions.
Ribosomal protein L-30 is a part of 50S ribosomal large subunit, synthesized in chloroplast. Schmidt et al. (1983) Sites of synthesis of chloroplast ribosomal proteins in Chlamydomonas. J Cell Biol 96(5):1451-1463