Plant/Algal cell antibodies / Photosynthesis / TOC/TIC

product information

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]

psTOC75; Predicted POTRA Domain #3; Expressed and purified in E. coli using the Impact System from NEB. Peptide confirmed by MALDI. Q43715

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Western blot (WB), flow cytometry (FACS), immunolocalization (IL)

AS08 345 | anti- Toc75 | Protein TOC75-3, chloroplastic, POTRA domain 1 rabbit antibodies

to be added when available

application information

1: 2000 - 1: 25 000, 1: 100 000 (WB), 1: 500 (FACS). 1: 100 (IL)

88 | 75 kDa (ocassionally a processing intermediate at 78 kDa is observed)

Arabidopsis thaliana, Pisum sativum, Spinacia oleracea, some  cross-reactivity was observed for cyanobacteria including:  Synechocystis, Synechococcus and Thermosynechococcus sp.

Ricinus communis, Vitis vinifera, Populus trichocarpa

no confirmed exceptions from predicted reactivity known in the moment

antibody detects Toc75 POTRA domain 3 as purified protein, in chloroplast fraction and in crude envelope fraction

to be added when available, antibody released February 2010

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 3 antibodies AS08 351 (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 (Millipore). Exposure time was 600 seconds.

Ashita Dave and Barry D. Bruce (UTK)