PC | Plastocyanin

Application example 

Thylakoid membranes (10 µg of total chlorophyll) extracted freshly from Hordeum vulgareleaves 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. 

Reactant: Malus domestica (Apple)

Application: Western Blotting

Pudmed ID: 23886449

Journal: Plant Methods

Figure Number: 2C

Published Date: 2013-07-28

First Author: Sikorskaite, S., Rajamäki, M. L., et al.

Impact Factor: 5.139

Analysis of protein fractions from different steps of nuclei isolation. The protein fractions obtained following the main steps of the procedure used for isolation of nuclei from leaves of apple are shown. Equal amounts of proteins (4 ?g) from each fraction were loaded on the gel. A) Lumenal-binding protein 2 (BiP2), B) histone H3 and C) plastocyanin (PC) were detected with specific antibodies by western blot analysis. D) Coomassie blue staining of the fractionated proteins separated by polyacrylamide gel electrophoresis. Lane 1: crude extract of proteins from homogenized leaf tissue; lane 2: resuspended pellet of the whole cell lysate obtained following treatment with Triton X-100 and centrifugation; lane 3: nuclei collected from the 60% Percoll layer (see nuclear fraction in Figure 1B); lane 4: the interface fraction of 60% Percoll and 2.5 M sucrose layers in the density gradient containing chloroplasts and unbroken cells (see 60% P/2.5 M S interface in Figure 1B); lane 5: sucrose layer of the density gradient (see 2.5 M sucrose layer in Figure 1B); lane 6: nuclei purified by centrifugation on a 35% Percoll cushion.

Reactant: Pisum sativum (Pea)

Application: Western Blotting

Pudmed ID: 33445673

Journal: Int J Mol Sci

Figure Number: 6A

Published Date: 2021-01-12

First Author: Tokarz, K. M., Weso?owski, W., et al.

Impact Factor: 5.542

Content of (a) plastocyanin (PC) and (b) large subunit of RuBisCo (RbcL) of grass pea leaf and stem photosynthetic apparatus under salinity; content of proteins expressed as relative units [RU]; different letters—statistically significant differences within each organ (leaf-uppercase, stem-lowercase) at p ? 0.05; (n = 3).