What could be the possible reasons behind low enrichment of isolated microvillous membrane of human placenta?

15 September 2022 0 9K Report

I have isolated microvillous (MVM) and basal plasma (BM) membranes from the human term placenta. After isolation I have checked the total protein content of isolated plasma membranes by BCA assay. Further to check the enrichment of isolated MVM I have performed alkaline phosphatase assay. For this I have added 1.25ug/5ul of MVM and 10ug/5ul of homogenate (Hom).

Enrichment Ratio= (Activity of MVM/µg of protein)/(Activity of HOM/µg of protein)

As I was getting low enrichment I have increased the concentration of MVM to 2.5ug/5ul by keeping the homogenate same. But still the enrichment is lower as the OD of homogenate is higher than usual. should I reduce the concentration of homogenate to 5ug/5ul? what could be the possible reasons for higher activity of alkaline phosphatase in homogenate?

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