eEF1a | Elongation factor 1-alpha
AS10 934 | clonality: polyclonal | host: rabbit | reactivity:A. thaliana, H.vulgare, Z.mays, O. europaea, P.patens
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1 : 1000 with standard ECL (WB)
|Expected | apparent MW||
Arabidopsis thaliana, Hordeum vulgare, Zea mays, Olea europaea, Physcomitrella patens
|Predicted reactivity||dicots including: Daucus carota, Glycine max, Nicotiana tabacum, Pisum sativum, Solanum lycopersicum, Vicia faba, monocots including: Oryza sativa, Triticum aestivum, trees: Malus domestica, other species: Lilium longiflorum, unicellular flagellate protists: Euglena gracilis|
|Not reactive in||
no confirmed exceptions from predicted reactivity known in the moment
to be added when available
to be added when available, antibody released in November 2012.
5 µg of total protein from Arabidopsis thaliana (A), Hordeum vulgare (B), Zea mays (C), Physcomitrella patens (P) extracted with Agrisera PEB buffer were separated on 4-12 % SDS-PAGE and blotted 70 min to PVDF. Blots were blocked with ECL Advance Blocking Reagent (GE Healthcare) for 1h at room temperature (RT) with agitation. Blot was incubated for 1 h with either rabbit pre-serum (1: 10 000), rabbit anti-eEF1alpha antibody (1: 10 000) and anti-eEF1 alpha serum incubated with eEF1alpha synthetic peptide (in 100 mM excess to saturate a signal from specific antibody) for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera ) diluted to 1:50 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 2 minutes.
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