HEN1 | HUA ENHANCER 1
AS15 3095 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 1000-1 : 5000 (WB)|
|Expected | apparent MW||
104.5 kDa | 105 kDa
|Confirmed reactivity||Arabidopsis thaliana|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
to be added when available, antibody released in January 2016.
50 µg of total protein from Arabidopsis thaliana whole vegetative rosette wild type Col-0 (a) and HEN1 overexpression mutant (b) extracted with extraction buffer (50 mM Tris pH 7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with Laemmli buffer at 95°C/5 min., were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Amersham ECL Prime and expose to Amersham Hyperfilms ECL for 20 seconds.
Courtesy of Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
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