DGAT2A | Acyl-CoA: Diacylglycerol acyltransferase
AS12 1874 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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Total proteins (containing 30 ug ) from Chlamydomonas reinhardtii cells grown for the indicated times in N-deprived medium extracted with lysis buffer (50 mM Tris-HCl, pH 6.8, containing 2% SDS and 10 mM EDTA and a protease inhibitor cocktail) were separated on 12 % SDS-PAGE and transferred onto a nitrocellose blot over night at 4°C. Blots were blocked with blocking buffer ( 5% (w/v) non-fat dry milk powder in TBS-T) for 2 hrs at room temperature (RT) with agitation. Blots were incubated in the primary antibody (CrTMDGAT2A) was used at a dilution of 1:1000 over night at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then whashed 5 times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:5000 in the same buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with Chemiluminescence detection kit according to the manufacturers instructions. An imaging system (ChemiDoc XRS; Bio-Rad) was used to quantitatively and qualitatively analyze protein blot. Exposure time was 30 seconds.
Courtesy Dr. Yantao Li, The University of Maryland, USA
Wase et al. (2015). Phenotypic screening identifies Brefeldin A/Ascotoxin as an inducer of lipid storage in the algae Chlamydomonas reinhardtii. Algal Research, Volume 11, September 2015, Pages 74–84.
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