Rnr3 | Ribonucleoside-diphosphate reductase large chain 2
AS09 574 | clonality: polyclonal | host: rabbit | reactivity: Saccharomyces cerevisiae
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|Recommended dilution||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 are currently known.|
|Additional information||Load per well was approx 3x10^6 cells (of a total extract).|
|Selected references||Lafuente-Barquero et al. (2017). The Smc5/6 complex regulates the yeast Mph1 helicase at RNA-DNA hybrid-mediated DNA damage. PLOS Genetics, December 27, 2017, doi.org/10.1371/journal.pgen.1007136
Schmidt et al. (2017). Alterations in cellular metabolism triggered by URA7 or GLN3 inactivation cause imbalanced dNTP pools and increased mutagenesis. Proc Natl Acad Sci U S A. 2017 May 30;114(22):E4442-E4451. doi: 10.1073/pnas.1618714114.
Graf et al. (2017). Telomere Length Determines TERRA and R-Loop Regulation through the Cell Cycle. Cell. 2017 Jun 29;170(1):72-85.e14. doi: 10.1016/j.cell.2017.06.006.
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.
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