Preparation of reaction solution: 0.1g G-250 was dissolved in 50 mL 90% ethanol, 100 mL of 85% phosphoric acid was added, and the volume was set to 1000 mL。
Assay: 12 μL Enzyme + 1.8 mL G-250 for 2 minutes, 595 nm colorimetric, blank (20 μL Buffer + 3 mL G-250).
You require a standard curve when using this method (Bradford). People often use bovine serum albumin for the standard protein. The standard curve consists of absorbance measurements made over a range of micrograms of the standard. Then you do the same test with your crude extract and use the standard curve to read the amount of protein from the absorbance.
Hi Kimbembe
Your question is not very clear, if you are trying to know the concentration of the enzyme then measure the the protein concentration this will give you an idea for how much protein you have used but remember this is not only your enzyme but with other proteins in your sample as you are using crude extract , the best way (if possible) purify your enzyme and measure protein concentration which mean a pure protein in this case.
Hope this helps
You require a standard curve when using this method (Bradford). People often use bovine serum albumin for the standard protein. The standard curve consists of absorbance measurements made over a range of micrograms of the standard. Then you do the same test with your crude extract and use the standard curve to read the amount of protein from the absorbance.
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