V-ATPase, a1 | vacuolar H+-ATPase subunit a isoform 1
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|Recommended dilution||1 : 1000 (WB)|
|Expected | apparent MW||
93 kDa (Arabidopsis thaliana)
|Confirmed reactivity||Arabidopsis thaliana|
|Predicted reactivity||Capsella rubella, Cucumis sativus, Erythranthe guttata , Glycine soja, Lupinus angustifolius, Morus notabilis, Phaseolus vulgaris , Vitis vinifera|
|Not reactive in||No confirmed exceptions from predicted reactivity are currently known.|
Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel.
To be added when available, antibody released in April 2017.
10 μg of microsomal membranes were isolated from 6-days-old etiolated (dark-grown) Arabidopsis thaliana seedlings with Extraction buffer (450mM sucrose, 50mM HEPES pH7.5, 5mM MgCl2, 1mM DTT, 1x protease inhibitor). Proteins were denaturated in SDS sample buffer for 5 min at 95°C. Proteins were separated on a 10% SDS-PAGE and blotted 1h to a nitrocellulose membrane using tank transfer. Blots were blocked with 4% dry milk in TBS-T for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody (#AS14 2822) at a dilution of 1:1000 in blocking buffer for 16h with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera #AS09 602) diluted to 1:25 000 in blocking buffer for 2h at RT with agitation. The blot was washed as above and developed for 3 min with ECL according to the manufacturer's instructions peqlab AceGlow Kit & INTAS digital imager. Exposure time was 3 min.
Courtesy of Fabian Fink, University Heidelberg, Germany
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