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CrPDAT1 | Phospholipid: diacylglycerol acyltransferase

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

CrPDAT1 | Phospholipid: diacylglycerol acyltransferase in the group Antibodies Plant/Algal  / Plant Developmental Biology / Lipid metabolism at Agrisera AB (Antibodies for research) (AS12 1875)
CrPDAT1 | Phospholipid: diacylglycerol acyltransferase



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Product Information

Immunogen recombinant CrPDAT1 without transmembrane domains, overexpressed in E.coli,
Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 350 ĩl
Reconstitution For reconstitution add 350 ĩl of sterile water
Storage Store lyophilized/reconstituted at -20°C; 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 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.

Additional information

Related products

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.
immunogen: recombinant CrPDAT1 without transmembrane domains, overexpressed in E.coli,
Reconstitution: For reconstitution add 350 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 350 ĩl
storage: Store lyophilized/reconstituted at -20°C; 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
calculated | apparent molecular mass [kDa]:

140 kDa

Confirmed reactivity: Chlamydomonas reinhardtii
predicted reactivity: Chlamydomonas reinhardtii
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer):

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: PDAT1 is an enzyme phospholipid: diacylglycerol acyltransferase, which catalyzes TAG (triglycerols) synthesis. It has a strong lipase activity with a broad substrate specificity.
All 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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