Dear scientific colleagues
I am performing lipase activity assay for Drosophila fat body (equivalent to human adipose tissue and liver). My issue is that my background control (sample homogenate which does not contain the lipase substrate) always show higher absorbance than my test samples which contains the lipase substrate. Since, the calculation involves subtracting background control absorbance from sample absorbance, the final absorbance is always negative which is erroneous. Can anyone please provide any suggestion on this?
P.S The fat body is highly enriched in triglycerides, so could there be a way to remove the inherent triglyceride/glycerol before proceeding with the assay.
Is the surface of the solution reflective? Is the control solution turbid (possibly from TGs)?
If possible, add the same amount of buffer (without lipase enzyme) to the control sample, as the buffer likely contains surfactants which change the surface characteristics of the fluid and/or solubilizes the fats making it less turbid
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