ABI5 | Abscisic acid insensitive 5
AS12 1863 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Expected | apparent MW||
|Predicted reactivity||Brassica napus, Populus trichocarpa|
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
|Additional information||ABI5 protein is present in very low levels therefore specific material should be used for analysis as well as most sensitive detection reagents.|
to be added when available antibody released in May 2015.
2, 5 and 10 ug of total protein (run on separate lanes) extracted from the Columbia ecotype (Arabidopsis thaliana) seeds using Acetone extraction was separated using the Bolt® Bis-Tris Plus Gel system on a 4-12% gradient SDS-PAGE gel, blotted using the turbo-blot system (BIO-RAD) to transfer onto a PVDF membrane (7min). SNAP-ID (Millipore) system was used for blocking and antibody labeling. Blocking occured for 30 minutes (no agitation, 0.05 % skim milk in dest. water). Primary antibody labeling was done for 10 minutes at 1:200 dilution. Followed by 3x10 ml washes (PBST). Then blotted with secondary antibody (anti-rabbit IgG HRP conjugated from Agrisera AS09 602) for 10 min (1:1000 dilution) followed by three washes.Blot was developed using ECL as per manufacturer’s instructions. Exposure time was 2 min.
Courtesy of Nay Chi, Pogson Lab, Australia
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