STM | Homeobox protein SHOOT MERISTEMLESS
AS11 1764 | Clonality: Polyclonal | Host: Rabbit | predicted Reactivity: A. thaliana
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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
Homeobox protein SHOOT MERISTEMLESS is a protein required for shoot apical meristem (SAM) formation during embryogenesis.