CrPDAT1 | Phospholipid: diacylglycerol acyltransferase

Product no: AS12 1875

AS12 1875  | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Chlamydomonas reinhardtii

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  • Product Info
  • Immunogen: recombinant CrPDAT1 without transmembrane domains, overexpressed in E.coli,
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Serum
    Format: Lyophilized
    Quantity: 350 l
    Reconstitution: For reconstitution add 350 l of sterile water
    Storage: Store lyophilized/reconstituted at -20C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
    Tested applications: Western blot (WB)
    Recommended dilution: 1 : 250 (WB) load per well up to 30 ug
    Expected | apparent MW:

    140 kDa

  • Reactivity
  • Confirmed reactivity: Chlamydomonas reinhardtii
    Predicted reactivity: Chlamydomonas reinhardtii
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • application example

    western blot using anti-PADT antibody

    Total proteins (containing 2.5 to 30 ug ) from Chlamydomonas cells 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 10 % 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. Blot was incubated in the primary antibody (∆TMCrPDAT) at a dilution of 1:250 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, from Bio-Rad) 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 of Dr. Kangsup Yoon, Arizona State University.

  • Background
  • Background: PDAT1 is an enzyme phospholipid: diacylglycerol acyltransferase, which catalyzes TAG (triglycerols) synthesis. It has a strong lipase activity with a broad substrate specificity.
  • Product Citations
  • Selected references: Yoon et al (2012). Phospholipid:Diacylglycerol Acyltransferase Is a Multifunctional Enzyme Involved in Membrane Lipid Turnover and Degradation While Synthesizing Triacylglycerol in the Unicellular Green Microalga Chlamydomonas reinhardtii. Plant Cell, Oct 2012.
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