phytase enzyme assay - (Jun/24/2008 )

hi...does anyone here know a colorimetric method to assay phytase activity? or a colorimetric method to quantify phosphate as that will be the end product released by phytate hydrolisis by phytase. i cant seem to find a method that works!! i've been having so much problem with the std curve as well

-khommy-

QUOTE (khommy @ Jun 24 2008, 08:24 AM)

I have successfully adapted the colorimetric inorganic phosphate assay of Murphy and Riley (1962) to a 96-well plate format. This is an ammonium molybdate reduction assay. I can post my protocol if you think you would find that useful. I have had no problems generating a nice linear std curve from 0.2 to 1.0 ug atomic phosphorus.

-jeffl-

QUOTE (jeffl @ Jun 25 2008, 01:56 AM)

QUOTE (khommy @ Jun 24 2008, 08:24 AM)

my god jeffl...u're a real life-saver! yes please, can u post ur protocol up here? I'll be forever in your debt. I've been on this for the past one yr...tried 3 differrent methods. They all worked initially but somewhere along the way sth always went wrong. I couldn't reproduce the same results for similar expmnts when tried at different times.

-khommy-

I've attached a copy of the protocol I recently developed. This particular assay was the basis of a large part of my advisor's PhD work so I can assure you it can give reproducible results. It has worked well for me so far. Let me know how it works out for you.

-transient-

thanks a bunch! will let u know how it goes soon!

-khommy-

Hi, do u have any protocols on tyrosinase assay? I need protocols on mushroom tyrosinase assay as well as cellular tyrosinase assay.

Hope can get some help. I am so lost in this enzymatic assay.

-sasoriza-

worthington has an assay for polyphenol oxidase (tyrosinase):

tyrosinase assay

-mdfenko-