Plant/Algal cell antibodies / Oryza sativa

product information

PGRL1 is a transmembrane protein present in thylakoids of higher plants and algae. Arabidopsis plants lacking PGRL1 show perturbation of cyclic electron transport (CEF) around photosystem I (PSI), similar to PGR5-deficient plants. PGRL1 has been shown to interact physically with PGR5 and associate with PSI (DalCorso et al., 2008). In Chlamydomonas reinhardtii, PGRL1 is part of a protein supercomplex, composed of PSI with its own light-harvesting complex (LHCI), the photosystem II light-harvesting complex (LHCII), the cytochrome b6/f complex (Cyt b6f), ferredoxin (Fd)-NADPH oxidoreductase (FNR), responsible of the energy balance of the two photosystems and of the switch between thylakoid linear and cyclic electron transport (Iwai et al., 2010). Synonymes:

the N-terminal loop (aa 61 to 200; N-PGRL1) of PGRL1A (At4g22890). The N-PGRL1 protein was purified by virtue of its His-Tag using Ni-NTA Agarose (QIAGEN).

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)

collection of antibodies to photosynthetic proteins

to be added when available

application information

1 : 3000 with standard ECL (WB)

29 | 29 kDa (Arabidopsis thaliana, Spinacia oleracea)

Arabidopsis thaliana, Spinacia oleracea

dicots:Glycine max, Ricinus communis, Vitis vinifera, trees: Populus trichocarpa, monocots: Oryza sativa, Zea mays

Physcomitrella patens, Chlamydomonas reinhardtii

In Arabidopsis thaliana samples cross-reactivity in the region of 55-60 kDa can be detected.

DalCorso et al., 2008. A complex containing PGRL1 and PGR5 is involved in the switch between linear and cyclic electron flow in Arabidopsis. Cell, 132: 273-285. Iwai et al., 2010. Isolation of the elusive supercomplex that drives cyclic electron flow in photosynthesis. Nature, 7292:1210-3.

application example

5 µg of chlorophyl a/lane from Arabidopsis thaliana wild-type thylakoids (1), Arabidopsis thaliana pgr5 mutant thylakoids (2), Arabidopsis thaliana double mutant thylakoids (3), were separated on  12% SDS-PAGE and blotted 1h to nitrocellulose membrane.Blots were blocked immediately following transfer with 5% dry milk in 1x TBS together with 0.1% Tween-20 (1x TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1:3000 for 2h at room temperature with agitation. The antibody solution was decanted and the blot washed three times, 10 min each, with 1x TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit horseradish peroxidase conjugated, from Pierce) diluted to 1:50.000 for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL detection reagent according to the manufacturer’s instructions. Exposure time was 5 min. 

Courtesy Paolo Pesaresi, Alexander Hertle, Roberto Barbato, Dario Leister, Ludwig-Maximilians-University Munich