Rnr1 | Ribonucleoside-diphosphate reductase large subunit
AS16 3639 | clonality: polyclonal | host: rabbit | reactivity: Saccharomyces cerevisiae
Replacement antibody for AS09 576.
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|Recommended dilution||1 : 5 000-1 : 10 000 (WB)|
|Expected | apparent MW||
99.56 | 100 kDa
|Confirmed reactivity||Saccharomyces cerevisiae|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Selected references||To be added when available, antibody released in March 2016.|
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
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