PsaC | PSI-C core subunit of photosystem I (PsaC antibody + PsaC protein positive control )
AS04 042P | clonality: polyclonal | host: rabbit | reactivity: [global antibody] for higher plants, algae, cyanobacteria
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|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana, Chlamydomonas reinhardtii, Cyanophora paradoxa, Heterosigma akashiwo, Emiliania huxleyi, Euglena gracilis, Gonyaulax polyedra, Horderum vulgare, Micromonas pusilla, Porphyra sp. Spinacia oleracea, Synechococcus PCC 7942, Thalassiosira pseudonan|
Algae, Cyanobacteria, Glycine max, Nicotiana tabacum, Physcomitrella patens, Prochlorococcus sp. (surface and a deep water ecotype), Spinacia oleracea
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
In some species minor cross reactions with some larger proteins are seen. These may contain related iron-sulfur binding motifs. Therefore size verification of the reacting band is required. Due to the small size of the protein, care should be taken to differentiate between chemiluminescent signal from PsaC and non-specific signals from chlotophylls or lipids if pigment is retained near the bottom of the blot.
Protein standard: use a load of 2 µl per well with ECL detection system and 4 µl per well with alkaline phospatase.
|Selected references||Ifuku et al. (2005). PsbP protein, but not PsbQ protein, is essential for the regulation and stabilization of photosystem II in higher plants. Plant Physiol. 3:1175-1184.
Oesterhelt et al (2007). Regulation of photosynthesis in the unicellular acidophilic red alga Galdieria sulphuraria. Plant J.3:500511.
2 µg of total protein from (1) Horderum vulgare leaf extracted with PEB (AS08 300), (2) Chlamydomonas reinhardtii total cell extracted with PEB (AS08 300), (3) Synechococcus sp. 7942 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: 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) diluted to 1:10 000 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).
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