PsbB | CP47 protein of PSII
AS04 038 | Clonality: Polyclonal | Host: Rabbit | Reactivity: [global antibody] for higher plants, Physcomitrella patens, algae, cyanobacteria, diatoms
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KLH-conjugated synthetic peptide derived from available plant, algal and cyanobacterial PsbB sequences including Arabidopsis thaliana AtCg00680, Hordeum vulgare P10900, Oryza sativa P0C364, Synechocystis PCC 6803 P05429
2 µg of total protein from Arabidopsis thaliana leaf (1), Horderum vulgare (2), Chlamydomonas reinhardtii total cell (3) Synechococcus sp. 7942 total cell (4), Anabaena sp. total cell (5), were extracted with PEB (AS08 300) and separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% blocking reagent 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: 50 000 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, recommended secondary antibody AS09 602) diluted to 1:50 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescence 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).
2.0 µg of chlorophyll from Pisum sativum chloroplasts and from Zea mays, Echinochloa crus-galli, Panicum miliaceum mesophyll and bundle sheath chloroplasts extracted with 0.4 M sorbitol, 50 mM Hepes NaOH, pH 7.8, 10 mM NaCl, 5 mM MgCl2 and 2 mM EDTA. Samples were denatured with Laemmli buffer at 75°C for 5 min and were separated on 12% SDS-PAGE and blotted 30 min to PVDF using wet transfer. Blot was blocked with 5% milk in TBS for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody AS04 038 at a dilution of 1: 2000 overnight at 4°C with agitation in 1% milk in TBS-T. The antibody solution was decanted and the blot was washed 4 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:25 000 in 1% milk in TBS-T for 1h at RT with agitation. The blot was washed 5 times for 5 min in TBS-T and 2 times for 5 min in TBS, and developed for 1 min with 1.25 mM luminol, 0.198 mM coumaric acid and 0.009% H2O2 in 0.1 M Tris- HCl, pH 8.5. Exposure time in ChemiDoc System was 122 seconds.
Courtesy Dr. Wioleta Wasilewska-Dębowska, Warsaw University, Poland
This product can be sold containing ProClin if requested.
This product can be sold containing ProClin if requested
in bis-tris gel systems PsbB protein migrates between 40-45 kDa
PsbB (CP47) is a chlorophyll-binding protein located in the membrane, where it serves as the core antenna of Photosystem II.
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