GPA1 | Guanine nucleotide-binding protein subunit alpha 1
AS12 2370 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana
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1 : 4000 with ECL (WB)
|Expected | apparent MW||
|Predicted reactivity||Brassica napus, Capsicum annuum , Cynara cardunculus var. scolymus, Eschscholzia californica, Glycine max, Gossypium hirsutum, Hordeum vulgare, Lupinus luteus, Medicago truncatula, Morus notabilis, Nicotiana benthamiana, Nicotiana tabacum, Oryza sativa, Pisum sativum, Phaseolus vulgaris, Populus trichocarpa, Ricinus communis , Solanum lycopersicum, Solanum tuberosum, Sorghum bicolor, Spinacia oleracea, Theobroma cacao, Triticum aestivum, Zea mays, Zostera marina, Vitis vinifera|
|Not reactive in||
No confirmed exceptions from predicted reactivity are currently known
The nature of a 37 kDa cross-reacting band is not known.
To be added when available, antibody released in April 2016.
25 µg of total protein from the indicated Arabidopsis thaliana wilde type and gpa1-3 mutant were extracted with the extraction buffer (250 mM 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 denatured with SDS loading dye (50mM Tris-HCl pH6.8, 100 mM DTT, 2%SDS, 10% glycerol, 0.025% bromophenol blue) at 70°C for 2-5 min were 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 10% skimmed milk powder for 2h and 20 min. at room temperature (RT) with agitation. Blot was incubated in the primary antibody 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 The secondary antibody (Agrisera Goat anti-rabbit IgG (H&L) HRP conjugate, AS09 602, 0.91 µg/µl) was diluted 1:5000 in the same solution and incubated on the membrane at room temperature for 2h. The membrane was then washed 5 times 15min with TBS-T (no milk powder) and the blot was developed using Super Signal West Pico and Femto (mixed 1:1) Chemiluminescent Substrate (Thermo Scientific). Exposure time was: 10 minutes.
Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany
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