Plant/Algal cell biology / Compartment markers / Chloroplast

product information

Antibody replaced by a new anti-Tic40 AS10 709

Tic40 is a component of the inner envelope membrane import complex (TIC) of plant chloroplasts. Tic40 has been proposed to function as a co-chaperone in the stromal chaperone complex that facilitates protein translocation across the inner membrane. Tic40 can be used as a cellular [compartment marker] for chloroplast inner envelope membrane.

KLH-conjugated synthetic peptide derived from available plant sequences of Tic40 including Arabidopsis thaliana At5g16620

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)

AS06 150 | anti-Toc75

AS08 293 | anti-Tic110

cellular [complartment marker] of chloroplast membrane

application information

1 : 5000 with standard ECL (WB)

48 | 45 kDa (Arabidopsis thaliana)

Arabidosps thaliana, Cucurbita pepo, Pisum sativum

Oryza sativa, Ricinus communis, Physcomitrella patens

Spinacia oleracea

to be added when available

not available at the moment

application example

10 µg of total protein from (1) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (2) chloroplast envelopes, 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-Tic40 (AS06 149, 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 (240s, standard sensitivity). The Tic40 signal obtained at ~45 kDa is completely quenched in a peptide-competition assay with the Tic40-peptide used for immunization. The signal at ~27 kda is not fully quenched and arises from a thylakoid-bound cross-reaction.