Dear all,

I am conducting an in-vitro anti-inflammatory assay, specifically protein denaturation assay by using bovine serum albumin. However, during the experiment using Aspirin as a control/drug reference, there was no reaction and the mixture only remained colourless. Plus, the absorbance reading at 660 nm also showed negative or zero. May I know any suggestion or solution behind this matter?

The protocols are as follows;

1) 0.05 ml of sample + 0.45 ml of 1% BSA

2) Incubate at 37 degrees Celcius for 20 minutes

3) Heat at 60 degrees Celcius for 3 minutes

4) Let to cool

5) Add 2.5 ml PBS

6) Measure the absorbance at 660 nm

I appreciate any help you can provide. Thank you in advance.