which media to be used, to make1.0 mM pNPGal solution?
which media to be used, to 100 μL of enzyme preparation ?
which media to be used, to make standard solution of p-nitrophenol (pNP)?
Should I use sodium acetate buffer to make all solutions or deionized water?
Na2CO3 used to stop the reaction in sample SOLUTIONS, so DO the same Na2CO3 to be used along with STANDARD pNP DILUTONS?
Hello,
1. pNPGal solution in deionized water
2. enzyme preparation in cold deionized water
3. 200 mM Sodium Acetate Buffer, w/w 0.25% pH 4.0 for reaction
4. standard solution of p-nitrophenol in deionized water
Na2CO3 stopping the reaction by increasing the pH of the reaction mixture.
Good luck!
Several papers used phosphate buffer 50mM or 20mM pH (6.8-6.9) to dissolve pNPG and enzyme DOI: 10.1155/2012/281078 or DOI: 10.1111/jam.12998
Hi Adarsh,
You can check 10.1074/jbc.M109.060145 for α-galactosidase assay. Good luck!
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