HY5 | Protein long hypocotyl 5
AS12 1867 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Recommended dilution||1: 1000 (WB)|
|Expected | apparent MW||
|Predicted reactivity||Brassica pekinensis|
|Not reactive in||
Oryza sativa, Pisum sativum, Populus sp. , Solanum lycopersicum
This antibody detects well recombinant HY5 in Nicotiana benthamiana (image below) while a signal in endogenous total cell extract of Arabidopsis thaliana has not been optimised in applied conditions (as below).
to be added when available, antibody released in April 2014.
20 ug of total protein from control (1), 35S::YFP-HY5-HA (2, red arrow), 35S::YFP-HY5-HA + 35S::CFP-X protein (green arrow), were separated on 12 % SDS-PAGE using tank transfer and blotted 1 h to PVDF (Biorad). Blots were blocked with 5 % skim milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HY5 antibody (second panel from the left) at a dilution of 1: 1000 for 1h at RT with agitation. 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 PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG, HRP conjugated from Agrisera, AS09 602), diluted to 1: 10 000 for 1h at RT with agitation. The blot was washed as above and developed for 5 min. with ECL according to the manufacturer's instructions. Exposure time was 60 seconds.
Courtesy of Dr. Seok Keun Cho, University of Copenhagen, Danmark
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