AGB1 | Guanine nucleotide-binding protein beta 1
AS16 3937 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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|Recommended dilution||1 : 5000 (WB)|
|Expected | apparent MW||41 kDa | 35 (37 kDa in 10 % gel) kDa|
|Confirmed reactivity||Arabidopsis thaliana
|Predicted reactivity||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.|
|Additional information||10 % gel is recommended to use for a better protein resolution.|
To be added when available, antibody released in March 2017.
10 µl of protein samples from Arabidopsis thaliana were separated on 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: 5000 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. The 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 and imaged with a BioRad Chemi Doc Touch system. Exposure time was: 10 minutes.
Total protein from the indicated Arabidopsis thaliana lines was extracted with extraction buffer CE ( 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). Protein concentration was measured with a Bradford assay and adjusted to 1mg/ml. Samples were denatured with SDS loading dye (50mM Tris-HCl pH6.8, 100mM DTT, 2%SDS, 10% glycerol, 0.025% bromophenol blue) at 70°C for 2-5 min.
Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany
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