One of my tests in my PhD research is estimation of catalase enzyme in liver of Rat, to see if there are any change in the level of this enzyme, after treatment of rat by sweeteners?
The isolation buffer for catalase consisted of 120 mM KCl, 2 mM K2CO3, 10 mM HEPES, and 1 mM EDTA; pH of 7.2 was adjusted with KOH.
We can use both Tris- Hcl and phosphate buffer for Catalase estimation
Prepare 10 % (w/v) liver homogenate in 50 mM ice cold phosphate buffer (pH 7- 7.2), centrifuge at 10,000g for 10 min and supernatant can be used to estimate enzyme activity
Prepare A 10% w/v homogenate in 50 mM phosphate buffer (pH 7.4) followed by cold centrifugation at 10,000 rpm at 4 °C for 20 min the post mitochondrial supernant (PMS) can be used for estimation of catalase and other antioxidant enzymes.
In my lab, we use KCl-histidine in a ratio 1:10 w/v. Thereafter, for CAT activity assay, you can use 50 mM phosphate buffer (pH 7.4).
I would like to Thank all researchers for there answer
I will use the buffer that are present in my lab.
thank you again
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