You could try this kit (link provided below). It can be adapted for cells or tissue samples by solubulizing samples in 5% Triton X-100.

http://www.bioassaysys.com/file_dir/ETGA.pdf

Hi Anuradha, Looks like a simple and easy method. You mention that it can be adapted for tissues. Have you any references to such protocols? I'm interested in perhaps using this for estimating fat content in fish liver.

Bjorn

I have used this kit in my samples and it works. I am only in the process of publishing the results.

Thank you, Anuradha, for your advise, we'll try that!

Hi, Anuradha,

would you be so nice to provide us with some details on the protocol? For example, we saw in other protocols that samples should be heated with 5% Triton. Is that necessary with your protocol? How much Triton/mg tissue do you use?

We really appreciate your help, thank you very much!

Antje

Hi Antje, I use isopropilic alcohol and then a commercial kit from Bioclin (Brazilian company) to measure TG. Its a kit for dosing blood TG. The protocol is attached, in portuguese, google translator may help you. It s a quite simple and fast protocol. Good luck!

Hi,

You can extract liver total lipids using Folch method, then you evaporate an aliquot of your lipid extarct by nitrogen flow to eliminate chloroforme. The dry extract must be dissolved in ethanol and then you can use a commercial TG Kit to dose TG level.

Good luck

I have used C:M mixture method to extract the cellular TG and used Spectrometer for the detection. You can use commercially available kit. However, I would suggest you to use TG colorimetric assay kit from Cayman. As they provide triglyceride standard along with it. https://www.caymanchem.com/pdfs/10010303.pdf

If you want to measure Tg in liver, you don't need to pulverise in nitrogen, simply put your frozen sample of liver in isopropanol (50mg/ml) and homogenize it by ultraturrax. Centrifuge at 4°C at 10000 rpm x 15min and take the supernatant full of Tg. You may use any colorimetric assay for Tg. Glycerol contamination in isopropanol extract is really little , but you may check that by not adding lipoprotein lipase if the kit allow that.

We usually extract the lipids from liver samples using hexan/isopropanol (3:2) over night. Afterwards aliquots of the extracts are dried and then the lipids are resolved in Triton/Chloroform. After drying we use a commercially available kit from Diagnostic Systems for spectrophotometric quantification of TG. We already published the protocol in several publications.

We measured the TG content in liver samples (which flash frozen in liquid nitrogen and kept in -80) spectrophotometrically by using the commercial TG kits (Sigma, ABCAM). we powdered liver samples, weighted about 40-60 mg of powder per tube, this part have to be done in cold (dry ice, liquid nitrogen), then add KOH, MG2SO4, reagents etc, just follow instruction. You can calculate your results as mg TG per g liver, mg TG per g liver protein or in whole liver, etc.

http://www.bio-protocol.org/wenzhang.aspx?id=223#.U1gUYaK83gs protocol for TG in liver

Dear Dr. Roberto Conti,

        Can you send me the exact protocol of Triacylglycerol estimation in liver homogenate using a spectrophotometric technique (Not necessary using a chemical kit). If you have a refined technical protocol, kindly forward it to me via the research gate or e-mail : [email protected]. Thanks for your help.

Lieber Dr. Alexandra,

        Danke fur der information bezuglich 'Triglycerid' ansicht im leber. Diese information wurde hilfe im unser experimentale prozedur. Aber ob ihnen habe ein laboratorie protokol betreffend diese technik mit benutzend der spektrofotometer bitte einschicken durch e-mail : [email protected].

Mince and homogenize tissues. Add 5%NP-40 and boil for 2min..cool and then boil again...cool..centrifuge..colllect supernatant. dilute accordingly and use a Sigma triglyceride detection kit. http://www.sigmaaldrich.com/catalog/product/sigma/tr0100?lang=en®ion=US

Infinity Triglyceride Reagent (Thermo) is probably the cheapest, and here is an easy protocol for extraction https://dgap.bu.edu/docs/LiverTG-protocol.pdf. Make a glycerol standard on your own, or buy one (glycerol or TG).