PIF4 | Phytochrome interacting factor 4 (goat antibody)
AS16 3955 | Clonality: Polyclonal | Host: Goat | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||48.3 | 60 kDa|
|Confirmed reactivity||Arabidopsis thaliana|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
|Additional information||Material used need to be up to 8 days old as detection in older rosette leaf may fail.|
to be added when available, antibody released in February 2017.
30 ul of total protein from Arabidopsis thaliana grown in 12 hour light/ 12 hour dark growth conditions and 6-day-old seedling samples taken at the end of the day (ZT12) were extracted with 100 mM MOPS, pH 7.6, 100 mM NaCl, 10% glycerol, 40 mM 2-mercaptoethanol, 5% SDS, 1X protease inhibitor cocktail from Roche, 2 mM PMSF. 80 µl buffer were added to about 100 µl grinded powder, then immediately heated at 70ºC for 10 minutes and separated on 8 % Bis-Tris SDS-PAGE and blotted 1h to PVDF using semi-dry or tank transfer. Blots were blocked with 5 % non-fat milk in TBST for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for over night at 4ªC with agitation in TBS-T. 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-goat 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 minutes with Thermo DURA (Thermofisher). Exposure time was 1 minute.
Courtesy of Dr. Bo Zhang, Umeå Plant Science Centre, Sweden
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