A12.2 | RNA polymerase I subunit (homolog of Pol II Rpb9)
AS07 225 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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10 µg of total protein from (1) Arabidopsis thaliana and (4) Oryza sativa leafs extracted with PEB (AS08 300), as well as (2) cytosolic and (3) nuclear fractions of Arbaidopsis thaliana leafs were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to nitrocellulose. Filters were blocked 1h with 2% low-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-A12.2 (AS07 255, 1:1000, 1h) and secondary anti-rabbit (1:20 000, 1 h) antibody (HRP conjugated) in TBS-T containing 2% low fat milk powder. Antibody incubations were followed by washings in TBS-T (15, +5, +5, +5 min). All steps were performed at RT with agitation. Signal was detected with chemiluminescent reagent using a Fuji LAS-3000 CCD (300s, standard sensitivity). The target A12.2 is specifically detected only in the nuclear extract of Arabidopsis thaliana (3).
This product has previously been labelled as anti-At3g25940 transcription factor S-II (TFIIS) domain-containing protein.
Protocol for isolation of cytosolic and nuclear fractions can be found here.
This antibody is specific for A12.2 subunit of RNA polymerase I but NOT RNA polymerase II or IV from Arabidopsis thaliana
RNA polymerase I is a nuclear located DNA-dependent enzyme involved in RNA elongation and regulation of transcription. In yeast subunit A12.2 has been described as homologous to the Rpb9 subunit from polymerase II.