PC | Plastocyanin

Purified native plastocyanin from Spinacia oleracea UniProt: P00289

10 kDa

Catalpa bungei, Dicots, Chlamydomonas reinhardtii, Nicotiana benthamiana, Physcomitrella patens, Ricinus communis, Solanum lycopersicum

Application example

Thylakoid membranes (10 µg of total chlorophyll) extracted freshly from Hordeum vulgare leaves with 100 mM HEPES-KOH (pH 7.5), 0.3 M sorbitol, 2 mM EDTA, and 1mM MgCl2 and denatured with a Laemmli buffer at 80°C for 5 min were separated on 12% SDS-PAGE and blotted 1 h to nitrocellulose (pore size of 0.2 um), using semi-dry transfer. Blot was blocked with 4% milk for 2 h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:3000 (PC and PetA, simultaneous western blot detection for both antibodies at the same time) for 1 h/RT with agitation in PBS-T. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:25000 in for 1 h/RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent according to manufactures recommendations. Exposure time was 30 seconds. Simultaneous western blot detection can be applied if MW of detected proteins differs in min. 20 kDa.

Courtesy Dr. Anja Liszkay, CNRS, France

10 µg of total protein from Arabidopsis thaliana (1),  Brassica juncea (2),  Zea mays (3), Oryza sativa (4),  Solamum lycopersicum (5), Nicotiana tabacum (6), Heliantus annuus (7) 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.

This antibody will also work well with HRP-conjugated secondary antibodies, as AS09 602. 

Cellular [compartment marker] of chloroplast thylakoid lumen

This product can be sold containing ProClin if requested.

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.

AS06 202 | Anti-cytc6 marker antibody of thylakoid lumen for Chlamydomonas reinhardtii, rabbit antibodies

Plant protein extraction buffer

Secondary antibodies

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. Alternative name: DNA-damage-repair/toleration protein DRT112.