Plant/Algal cell antibodies / Populus sp.

product information

PSII reaction centre components are  generating the redox potential required to drive highly oxidizing water splitting reaction. Four Mn atoms are present on a lumenal surface and form the catalyctic site          of the water-splitting reaction which is in close association with the 33 kDa (PsbO), 23 kDa (PsbP) and 17 kDa (PsbQ) extrinistic subunits of oxygen evolving complex OEC. A 33-kDa extrinsic protein is also termed the Mn-stabilizing protein (MSP), however recent evidences shown that it is C-terminal domain of PsbA (D1) protein which is involved in in the assembly and stabilization of the OEC.

native purified 33 kDa protein 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)

AS05 092 | anti-PsbO | 33 kDa of the oxygen evolving complex (OEC) of PSII

AS06 167 | anti-PsbP | 23 kDa protein of the oxygen evolving complex (OEC) of PSII

AS08 305 | anti-PsbP | 23 kDa protein of the oxygen evolving complex (OEC) of PSII

AS06 142-16 | anti-PsbQ | 16 kDa protein of the oxygen evolving complex (OEC) of PSII

total IgG fraction has been purified by 40% ammonium sulpgate precipitation followed by DEAE cellulose chromatography

application information

1:2000-1: 5000  with standard ECL (WB)

35 | 33 kDa

Arabidopsis thaliana , Hordeum vulgare, Pinus banksiana, Spinacia oleracea, Chlamydomonas reinhardtii, Synechococcus sp. PCC 7942

dicots including: Nicotiana tabacum, Solanum lycopersicum, trees: Populus trichocarpa

no confirmed exceptions from predicted reactivity known in the moment

to be added when available

Camm et al. (1987). Association of the 33 kDa extrinistic polypeptide (water-splitting) with PSII particles: immunochemical quantification of residual polypeptide after membrane extraction. Photos Res 13: 69-80.

application example

2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with PEB (AS08 300), (2) Hordeum vulgare leaf extracted with PEB (AS08 300), (3) Chlamydomonas reinhardtii total cell extracted with PEB (AS08 300), (4) Synechococcus sp. 7942 total cell extracted with PEB (AS08 300), (5) Anabaena sp. total cell extracted with PEB (AS08 300) were separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Abcam) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL Advance detection reagent according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad).