eEF1a | Elongation factor 1-alpha

KLH-conjugated synthetic peptide derived from known sequences of eEF1a including Arabidopsis thaliana EF-1-alpha 1P0DH99 AT1G07940  EF-1-alpha 2 Q8W4H7  AT1G07930, EF-1-alpha 3 Q0WL5 AT1G07920, EF-1-alpha 4 Q8GTY0 AT5G6039

Aapplication example

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.

This product can be sold containing ProClin if requested.

Following protein resolubilization with urea/thiourea/CHAPS signal strength of recognized band is decreased. Recommended protein extraction conditions are without urea.

EF1A (elongation factor 1-alpha) belongs to the GTP-binding elongation factor family and is localized in the cytoplasm. It is an essential enzyme in elongation phase of protein synthesis. There are four genes encoding EF1A in Arabidopsis, sequences of genes 1,2 and 3 are identical. Synonymes:eEF-1A.