RpoTm | DNA-directed mitochondrial RNA polymerase (RpoT1; mitochondrial NEP)
AS15 3078 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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2.5 ng (1), 5 ng (2), 10 ng (3) of recombinant Arabidopsis thaliana RPOTm and 40 µg of a total cell extract from Arabidopsis thaliana leaves (4) were separated on 12 % SDS-PAGE and blotted 1h to nitrocellulose using tank transfer. Blots were blocked with 5 % milk in TBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2 000 for 1h 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 5 min with ECL according to the manufacturer's instructions. Exposure time was 1 to 5 seconds.
Band at 110 kDa represents fully matured protein.
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