DRP5B | Dynamin related protein 5B
AS12 2634 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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1 : 5000 (WB)
|Expected | apparent MW||
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Glycine max, Manihot esculenta, Mesostigma viride, Oryza sativa, Physcomitrella patens, Sorghum bicolor, Zea mays, Vitis vinifera|
|Not reactive in||
no confirmed exceptions from predicted reactivity are currently known
most sensitive ECL reagents are necessary to obtain a reaction with this antibody
to be added when available, antibody released in May 2014.
Total proteins extracted from 5 mg leaf or flower bud of Arabidopsis thaliana tissue with SDS-loading buffer were separated on 10% SDS-PAGE using tank transfer and blotted for 1 h to nitrocellulose membrane. Blots were blocked with 2% non-fat dry milk for 1 h at room temperature (RT) with agitation. Blot was incubated in the primary antibody (affinity-purified DRP5B) at a dilution of 1:5 000 in TBS-T + 2% non-fat dry milk overnight at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horseradish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:5 000 in TBS-T + 2% non-fat dry milk for 2 hr at RT with agitation. The blot was washed as above and developed for 5 min with SuperSignal West Pico (Thermo Scientific) according to the manufacturer's instructions. Exposure time was 30 min. Additionally the same blot was washed and developed with SuperSignal West Femto (Thermo Scientific) for 1 s. Exposure time was 1 min. The latter helped to increase DRP5B detection with the affinity-purified DRP5B antibodies. The signal was not obtained from bud tissue, suggesting that expression level of DRP5B may be too low for detection (blot not included).
Courtesy of Deena Kadirjan-Kalbach and Katherine W. Osteryoung, Michigan State University, USA
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