What is the best way to purify lectin from mushrooms (PLEUROTUS OSTREATUS) using a lactosyl-sepharose 4B coloum?

16 April 2015 1 3K Report

i extracted using PBS pH 7.1 and the raw extracts lyse RBC when tested for agglutination using microwells. After passing the extract in the coloum, i eluted with 0.2M lactose in thesame buffer and dialysed but i cant see agglutinating activity when tested for using microwells plate. please is there any place i'm getting it wrong? or is there any other simpler method using lactose?

Ravi Kant Upadhyay

Use 0.e M lactose, and make the pH above 7.5, you will get your positive result. Reduce the time of treatment anddo it in cold at 4 degree temperature.

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