DISCONTINUED Rnr1 | Ribonucleoside-diphosphate reductase large subunit
AS16 3639 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Saccharomyces cerevisiae
Replaced by AS21 4608
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10 ul of total protein from a 1 x 108 cells/ml culture of Saccharomyces cerevisiae extracted with 20% TCA and re-suspended in 1X loading buffer (4X SDS loading buffer: 150 mM TrisHCl pH 7.0, 25% Glycerol, 12% SDS, 0.02% Bromophenol Blue, 5% B-mercaptoethanol) were separated on 7.5% SDS-PAGE and blotted to nitrocellulose membrane for 2h/60V using tank transfer. Blots were blocked (5% milk) for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1:10000 or 1:5000 o/n at 4°C. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:50 000 or 1: 75 000 in 5% milk for 1h at RT with agitation. The blot was washed as above and developed with Western Lightning ECL Pro (PerkinElmer). Exposure time was between 30 seconds and 1 minute depending on the dilutions.
Courtesy of Dr. Isaac Corcoles, NWCR, United Kingdom
Saccharomyces cerevisiae Rnr1 (EC=126.96.36.199) is an enzyme from ribonucleoside diphosphate reductase large chain family. Is localized to cytoplasm and provides precursors necessary for DNA synthesis. Alternative names: ribonucleotide reductase large subunit 1, ribonucleotide reductase R1 subunit 1
Corcoles-Saez et al. (2019). Functional link between mitochondria and Rnr3, the minor catalytic subunit of yeast ribonucleotide reductase. Microb Cell. 2019 May 20;6(6):286-294. doi: 10.15698/mic2019.06.680.
Sampaio-Marques et al. (2019). α-Synuclein toxicity in yeast and human cells is caused by cell cycle re-entry and autophagy degradation of ribonucleotide reductase 1. Aging Cell. 2019 Aug;18(4):e12922. doi: 10.1111/acel.12922.
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