Hi All,
I'm need to perform an IFNalpha ELISA on mouse plasma samples, however these samples are from mice that received a high cholesterol diet and therefore the plasma is turbid because of the lipids.
As this will most likely influence my ELISA results, I'm looking for a method to avoid this problem. For example, is there a way to get rid of the cholestrol/lipid without influencing the IFNa level in the sample so that I can still quite accurately measure it with ELISA?
I also though about CBA analysis instead of ELISA, but there are no kits available for mouse IFNa. Also I would not know whether it interferes with the fluoresence of the assay.
Can someone help me out?
Cheers
Bart
You may try the way of fasting your mice 4-5 hours prior to collecting blood samples, that is how I avoid the hyperlipidemia when I did the ELISA with high fat diet fed mouse model .
You may attempt to clarify the samples by centrifugation at high speed (> 10,000 rpm for 30 minutes) in a microfuge to microtubes (0.5 ml) and recovering the infranatant (large Lipoproteins floating) for ELISA analysis. If clarification is insufficient, you should use UC, but I think if the lipid components are chylomicrons and large VLDL, with this approach may be sufficient.
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