RpoB | RNA polymerase beta subunit (chloroplast) (maize)
AS15 2867A | Clonality: Polyclonal | Host: Rabbit | Reactiviy: Zea mays
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Stroma, thylakoid, nuclei and total protein from Zea mays were extracted with 50 mM HEPES-KOH pH 8, 330 mM sorbitol exaction buffer. Components were separated on 12% SDS-PAGE and blotted overnight to Nitrocellulose membrane using wet transfer. Blots were blocked with 5% non-fat milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 500 for 2h at RT 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 RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5min with chemiluminescent detection reagent of extreme low femtogram range. Exposure time was 120 seconds.
Courtesy of Jie Shen and Dr. Alice Barkan, University of Oregon, USA
This antibody is detecting recombinant RpoB.
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