Rnr3 | Ribonucleoside-diphosphate reductase large chain 2
AS09 574 | clonality: polyclonal | host: rabbit | reactivity: Saccharomyces cerevisiae
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1: 500-1:1000 (WB)
|Expected | apparent MW||
97.5 | 98 kDa
|Confirmed reactivity||Saccharomyces cerevisiae|
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
load per well was approx 3x10^6 cells (of a total extract)
|Selected references||Williams et al. (2017). The role of RNase H2 in processing ribonucleotides incorporated during DNA replication. DNA Repair (Amst). 2017 Mar 6. pii: S1568-7864(16)30431-1. doi: 10.1016/j.dnarep.2017.02.016.
Tumbale et al. (2013). Aprataxin resolves adenylated RNA-DNA junctions to maintain genome integrity. Nature. 2013 Dec 22. doi: 10.1038/nature12824.
Golla et al. (2013). Sen1p Contributes to Genomic Integrity by Regulating Expression of Ribonucleotide Reductase 1 (RNR1) in Saccharomyces cerevisiae. PloS One, May 31.
Tsaponina et al. (2011). Ixr1 Is Required for the Expression of the Ribonucleotide Reductase Rnr1 and Maintenance of dNTP Pools. PLoS Genetics.
10 μl total protein from 9.25 x 107 cells of Saccharomyces cerevisiae extracted with 20% TCA as described below were separated on 10% SDS-PAGE and blotted 1.5h (0.5 A) to a nitrocellulose membrane (Whatman PROTRAN BA 85, 0.45 μm). Blots were blocked with 5% non-fat dry milk in TBST for 1.5h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 200 overnight at 4C° with agitation. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBST at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, (AS09 602) diluted to 1:50 000 for 1h at RT with agitation. The blot was washed as above and developed for 3 min with SuperSignal West Pico Stable Solution (1856135 Thermo Scientific) and SuperSignal West Pico Luminol/Enhancer Solution (1856136 Thermo Scientific) mixed 1:1 rate according to the manufacturers instructions. Exposure time was 10 min.
Courtesy Dr. Andrei Chabes, Umeå University, Sweden