AGB1 | Guanine nucleotide-binding protein beta 1
AS16 3937 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 1000|
|Expected | apparent MW||41 kDa | 35 kDa|
|Predicted reactivity||Arabidopsis thaliana, Brasica sp., Cajanus cajan, Camelina sativa, Capsella rubella, Eutrema sp., Cicer arietinum, Gossypium sp., Medicago truncatula, Morus sp., Cajanus cajan, Pisum sativum, Sesamum indicum, Solanum sp., Tarenaya hassleriana, Theobroma cacao, Trifolium subterraneum
|Not reactive in||
No confirmed exceptions from predicted reactivity are currently known
To be added when available
To be added when available.
The samples extracted either directly with SDS loading buffer (50mM Tris-HCl pH6.8, 100mM DTT, 2%SDS, 10% glycerol, 0.025% bromophenol blue) and then denatured at 95°C for 5 min, or first extracted with CE buffer (250mM sucrose, 100mM HEPES-KOH pH 7.5, 5% glycerol, 1mM Na2MoO4 x 2H2O, 25mM NaF, 10mM EDTA, 1mM DTT, 0.5%Triton X-100, protease inhibitor cocktail) and then denatured with SDS loading buffer at 70°C for 5 min. 10 µl of proteins were loaded into each well and separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with Tris-buffered saline containing 0.05% Tween-20 (TBS-T) and 5% skimmed milk powder for 1h at room temperature (RT) with agitation. The blot was incubated in the primary antibodies indicated at a dilution of 1: 5 000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 5 times for 15 min in TBS-T with milk powder at RT with agitation. Blot was incubated in secondary antibody (Goat-Anti-Rabbit AP conjugate, Sigma) diluted 1:5000 for 2h at RT with agitation. The blot was washed as above with TBS-T without milk powder, equilibrated in AP buffer (100mM TRIS pH=9.5, 100mM NaCl, 50mM MgCl2) and then developed with BioRad Immunstar AP substrate, before exposure to a CEA RP NEW film for 20 s.
Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany
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