Plant/Algal cell biology / Compartment markers / Chloroplast

product information

Import of protein precurors into chloroplasts occurs via translocon complexes at the outer (Toc/TOC complex) and inner (Tic/TIC complex) envelope membrane. The major components of the Toc (TOC) complex in plants are Toc75, Toc34 and Toc159 (formerly Toc86). Toc75 (OEP75) forms the main import pore. It is an abundant protein in the outer envelope membrane of chloroplasts from expanding tissues but it is also found in plastids of roots, stems and flowers.

KLH-conjugated synthetic peptide conserved in available Toc75 (OPE75) sequences from monocots and dicots including Arabidopsis thaliana (Q9STE8). The Arabidopsis genome contains three Toc75-related sequences, termed atTOC75-I, atTOC75-III, and atTOC75-IV with atToc75-III being the most expressed one.

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)

AS08 292  |  anti-Toc75 (Pisum sativum)

AS07 239  |  anti-Toc159 (guinea pig antibody)

Toc75 can be used as a cellular [compartment marker] of chloroplast outer envelope membrane.

application information

1 : 1000 - 1: 5000 with standard ECL (WB)

89 kDa (with transit peptide) | 75 kDa (for Arabidopsis thaliana)

Arabidopsis thaliana, Oryza sativa, Pisum sativum

dicots including: Ricinus communis, Vitis vinifera and Populus sp., monocts including Zea mays, Physcomitrella patens

no confirmed exceptions from predicted reactivity known in the moment

to be added when available

to be added when available

application example

10 µg protein from (1) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (2) chloplast envelope membranes, and (3) thylakoids were separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-Toc75 (AS06 150, 1:1000, 1h) and secondary anti-rabbit (1:20000, 1 h) antibody (HRP conjugated, Abcam) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with standard ECL (Invitrogen) using a Fuji LAS-3000 CCD (300s, standard sensitivity).