Toc75 | Protein TOC75-3, chloroplastic, POTRA domain 1

Application example

580 ng of Chl of Pisum sativum plants (10 day old) (2, 4)  and  10 µg of combined envelopes of Pisum sativum 10 day-old (1,3) were separated on 15% SDS-PAGE and blotted 2h to PVDF. PVDF was blocked 1h with 3% non-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-Toc75 POTRA domain 1 antibodies AS08 345 (1:2000 and 1: 5000, 1h) and secondary donkey-anti-rabbit (1:20000, 1 h) antibody (HRP conjugated) in TBS-T containing 3% non fat milk powder. Antibody incubations were followed by washings in TBS-T. All steps were performed at RT with agitation. Blots were developed for 5 min with HRP substrate Peroxide solution & luminol detection reagent according to the manufacturers instructions. Exposure time was 600 seconds.

Courtesy of Ashita Dave and Barry D. Bruce (UTK), USA

OEP 75 or Toc75; Chloroplast outer envelope membrane protein from Pisum sativum (pea), Predicted to contain 3 POTRA domains at N-terminus. Believed to be the protein conducting channel of the Toc translocon and assembles as an 18 stranded ß-barrel (EMBO J. (1995) 14:11, 2436-2446); In Arabidopsis there are five members of this Family Toc75 (I-V), atToc75III is most closely related to psToc75. Additionally, it is structurally related to members of the bacterial surface antigen super-family including: OMA87; Outer membrane protein/protective antigen, (COG4775, COG4775) [Cell envelope biogenesis, outer membrane;  YaeT; outer membrane protein assembly complex, (TIGR03303);  FhaC; Hemolysin activation/secretion protein (COG2831) [Intracellular trafficking and secretion]