Quality Lipoproteins | Reliable Results
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FAQ

What happens to the lipoproteins during storage?

 

The lipid components of the lipoproteins are susceptible to oxidation. The buffer we use acts as a preservative that deters oxidation from metal ions.

Once oxidation has begun, there are many fast acting, short lived, reactive products generated that interact with the lipid and /or protein portion of the particle. Changes may occur over days to weeks regardless of the buffer used for storage.

The purified lipoproteins may begin to aggregate after storage. Rough handling (such as vortexing or rapid pipetting) will accelerate this process. After months of storage, precipitates may form and the concentration of lipoproteins in solution will decrease. Fluorescently labeled lipoproteins will lose fluorescence intensity after a several months of storage. They are also prone to aggregation (see above).

How is the concentration of the lipoprotein determined?

 

The protein concentration is determined by the Pierce 660 nm Protein Assay using bovine albumin / gamma globulin for the standard curve.

The protein concentration determined by another assay or using another standard curve may give different results.

What are the best handling techniques to prolong the shelf life of lipoproteins?

 

The products are provided in a buffer containing EDTA as a preservative and sealed under nitrogen gas when packaged.

  • To prolong the shelf life of lipoproteins use aseptic technique whenever removing an aliquot.
  • Handle the liquid gently (no vortexing) to prevent aggregation.
  • Do not freeze the products to avoid aggregation.
  • Do not allow fluorescent labeled lipoproteins to be exposure to light for prolonged time periods.

How do I detect the fluorescent lipoproteins by microscope?

 

You can detect the fluorescence using filter settings for the following:

Fluorescent label

Excitation/Emission (nm)

DiO (3,3'-dioctadecyloxacarbocyanine)

 

482/501

DiI (1,1'-dioctadecyl- 3,3,3',3'-tetramethylindocarbocyanine perchlorate)

549/565

 

Specific configurations are available from the equipment manufacturers:

Manufacturer

Link

Nikon

www.microscopyu.com/articles/fluorescence/filtercubes/green/tritchyq/tritchyqindex.html

Olympus

http://www.olympusmicro.com/primer/techniques/fluorescence/fluorotable1.html

Leica

http://www.leica-microsystems.com/science-lab/fluorescent-dyes/

How do I detect the fluorescent lipoproteins by flow cytometer?

 

DiO is excited at 488 nm and detected with FITC emission filters (eg.533nm/30 or 525nm).

DiI is excited at 488 nm and detected with PE emission filters (eg.585nm/42).

What is the cholesterol content of the Lipoprotein Depleted Fetal Bovine Serum?

 

We do not routinely assay for the cholesterol content of our Lipoprotein Depleted Fetal Bovine Serum. Our routine release testing includes the comparison of the starting serum with the depleted serum by agarose gel electrophoresis. We stain the gel with Sudan Black (a lipid stain) and confirm that the lipoproteins are depleted.

To check our depletion process, we have compared one preparation of the starting serum and depleted serum to verify that the cholesterol is indeed reduced. Using the HDL and LDL/VLDL Cholesterol Assay Kit from abcam (ab65390),we found a forty fold reduction.

Here are the results for that assay:

Sample Identification

Cholesterol (mg/mL)

Protein (mg/mL)

Fetal Bovine Serum

1.40

35.0

Lipoprotein Depleted Fetal Bovine Serum

0.04

37.5

Why does my Lipoprotein Depleted Fetal Bovine Serum look different from the picture online?

Our Fetal Bovine Lipoprotein Depleted Serum is packaged in a High-Density PolyEthylene (HDPE) bottle so it can withstand storage of -80 degrees centigrade for long periods of time. The bottle itself is slightly opaque. The picture on the website, however, is a completely thawed serum with a transparent bottle (the clear bottle is a bit more photogenic than the HDPE bottles).