For protein extraction i found a protocol that separate phosvitin from egg yolk using ultrafiltration trough 10 MWCO then uses the filtrate. But phosvitin molecular weight is 35 KDa. So how it is possible to collect phosvitin in the filtrate? Another protcol uses heating temperature to seperate phosvitin but i want to compare phosvitin stability before and after heat treatment. So heating during extraction is not a good option for me.
Any suggestion will be helpful.
Thanks.
Phosvitin was extracted from yolk granules using 10% NaCl or 10% (NH4)2SO4 (final concentration) and then treated with heat to precipitate the lipoproteins from the extracted solution. The optimal pH for the phosvitin extraction from yolk granules was determined, and the iron-binding ability of the extracted phosvitin (final product) was tested. Adding 10% (NH4)2SO4disrupted the granules, and the subsequent thermal treatment at 90°C for 1 h precipitated low density and high density lipoproteins, which enabled separation of phosvitin by centrifugation.
Rohit Kumar thanks for your answer but i want to compare the heat stability of the protein before and after treatment, so using the heat for extraction affect my results.
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