Plant/Algal cell biology / Compartment markers / Chloroplast

product information

Plastocyanin (PC) is a small Cu protein and a mobile electron carrier in the lumen of the thylkoids. PC interacts with the B/F complex and Photosystem I.

purified native plastocyanin from Spinacia oleracea

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), immunogold (IG)

AS06 202 | anti-cytc6 marker antibody of thylakoid lumen for Chlamydomonas reinhardtii

cellular [compartment marker] of chloroplast thylakoid lumen

application information

1:2000 with alkaline phosphatase (WB), 1: 100 (IG)

10 kDa

Arabidops thaliana, Brassica juncea, Heliantus annuus, Nicotiana tabacum, Oryza sativa, Pisum sativum, Spinacia oleracea, Solanum tuberosum, Zea mays

dicots Ricinus communis, Solanum lycopersicum, monocots including Hordeum vulgare, moss Physcomitrella patens

no confirmed exceptions from predicted reactivity known in the moment

plastocyanin runs abberant due to negative charge at 12-19 kDa on SDS-PAGE depending upon the system used. in 15 % gel the protein will run closer to its true MW than in 12 % gel. In some cases PC can be very acidic and run at twice of its MW.

PC1 runs closer to 14 kDa while PC2  runs closer to 19 kDa.  For good resolution adding fresh DTT to the sample buffer is recommended.
PC2 is generally more abundant and it increases with Cu feeding. PC1 is expressed first after etiolated seedlings are placed in the light.

Abdel-Ghany et al. (2005) Two P-type ATPases are required for copper delivery in Arabidopsis thaliana chloroplasts. Plant Cell 17 1233-1251

application example

10 µg of total protein from (1) Arabidopsis thaliana, (2) Brassica juncea, (3) Zea mays, (4) Oryza sativa, (5) Solamum lycopersicum, (6) Nicotiana tabacum, (7) Heliantus annuus were separated on  SDS-PAGE and blotted to nitrocellulose. Filters were probed with anti-PC antibody  (AS06 141, 1:2000). Signal was developed using alkaline phosphatase conjugated secondary antibody. Each sample was run in duplicate. Signal was developed using alkaline conjugated secondary antibody.