DGAT2A | Acyl-CoA: Diacylglycerol acyltransferase

AS12 1874 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii


product information
Background		DGAT2A (Acyl-CoA: Diacylglycerol acyltransferase) is an enzyme which exhibits transferase activity, transferring acyl groups. EC=2.7.8.2.
Immunogen		recombinant CrDGAT2A, overexpressed in E.coli, missing transmembrane domains, PID 536226, also annotated as DGTT1 UniProt: A8JGY1
Host		Rabbit
Clonality		Polyclonal
Clone
Purity		Serum
Format		Lyophilized
Quantity		100 µl
Reconstitution		For reconstitution add 100 µl of sterile water.
Storage		Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications		Western blot (WB)
Related products		Algal protein extraction buffer Secondary antibodies
Additional information

application information
Recommended dilution		1 : 1000 (WB)
Expected \| apparent MW		kDa
Confirmed reactivity		Chlamydomonas reinhardtii
Predicted reactivity		Chlamydomonas reinhardtii
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references		Liu et al. (2016). A simple and reproducible non-radiolabeled in vitro assay for recombinant acyltransferases involved in triacylglycerol biosynthesis. J Appl Phycol (2016). doi:10.1007/s10811-016-0949-6. 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.

application example

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 (Bio-Rad) 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

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