How do I label chylomicrons with a fluorescent probe (specific to chylomicrons)?

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No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

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Olivier Catrice

Nile Red ? It is a vital bright fuorescent lipid dye.

http://jcb.rupress.org/content/100/3/965.full.pdf

Feilong Sun

Thanks for your reply. But I am afraid Rile red can also stain the cell membrane when I use it for chylomicrons.

Just try and see.

Is it really a problem if the plasma membrane is also stained if you want to observe droplet within the cell ?

Plasma membrane has a morphology completely different from droplets, so it is easy to discriminate them by shape.

Thanks for your answer

Michael Maher

Is it the triglyceride- or phosopholipid-specific Nile Red that you are using?

Also ApoB48 is specific to Chylomicrons, so a labelled antibody against this marker may help you

Thanks Michael, Nile red, even phospholipid specific probe, can also label lipid droplets in cyto- plasma. As far as I know, apoB48 is also expressed in VLDL. Anyway, antibody targeting to apo-b48 can be a good choice to trace chylomicrons so far.