STM | Homeobox protein SHOOT MERISTEMLESS
AS11 1764 | clonality: polyclonal | host: rabbit | predicted reactivity: A. thaliana
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1 : 1000 with standard ECL (WB)
|Expected | apparent MW||
|Predicted reactivity||Brassica napus, Brassica oleracea|
|Not reactive in||
no confirmed exceptions from predicted reactivity are currently known
to be added when available
to be added when available, antibody released in June 2015
10 µg of total protein from Arabidopsis thaliana seedling tissue extracted with Exractions buffers 1 or 2 (see below) were separated on 10 % SDS-PAGE and blotted 1h to PVDF using semi-dry transfer. Blots were blocked with milk for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in milk overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed twice with milk, then washed once for 5 mins with PBS, 10 mins with 2% milk + PBST, 5 mins with PBST at RT with agitation. Blot was incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:75 000 in 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 variable.
1. 50mM tris-acetate pH 7.9 100mM potassium acetate 20% glycerol 1mM EDTA 1mM DTT 1x Roche Protease inhibitor cocktail
2. 100mM tris-cl pH 7.5 75mM Nacl 15mM EGTA 15mM MgCl2 1mM DTT 400 mM B glycophosphate 1x protease inhibitor cocktail NaFl, Na pyrophosphate, Na vanadate.
Courtesy Dr. Simon Scofield, Cardiff University, UK
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