Elip | Early light induced protein
AS06 147A | Clonality: Polyclonal | Host: Rabbit | Reactivity: Ch. reinhardtii, P. sativum, Vitis sp.
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20 kDa (Pisum sativum)
About 30 µg of total protein from Chlamydomonas reinhardtii, extracted with pre-cooled buffer (10% glycerol, 20 mM HEPES, 5 mM MgCl2, 2.5 mM EDTA, 10 mM KCl, 1 mM PMSF, 0,5% DTT; pH=7,4) and denatured with SDS/mercaptoethanol loading buffer at 95C for 5 min, were separated on 12% SDS-PAGE and blotted at 60V for 2,5h to nitrocellulose membrane using tank transfer. Blots were blocked with 0.2% Tween-20 in TBS (overnight, at 5OC) with agitation. Blot was incubated in the primary antibody at a dilution of 1:500 in TBS-T (0.05% Tween-20 in TBS) overnight at 5OC with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from goat) diluted 1:15 000 in TBS-T, for 2h at RT with agitation. The blot was washed as above and developed using *DAB/H2O2 mixture in TBS for approximately 5 min.
* DAB = 3,3'-Diaminobenzidine
Courtesy of Dr. Anna Aksmann, Department of Plant Physiology and Biotechnology, University of Gdańsk, Poland
Min. 30 µg of total protein has to be loaded per lane.
Early light-induced proteins (ELIPs) are light stress-induced proteins related to the chlorophyll a/b binding protein family from higher plants and green algae located in the thylakoid membranes and involved in photosynthesis.