We have performed an inhibition of egg albumin denaturation assay to access the efficacy of natural plant extract,

At 5 ml total volume; 0.2 ml of 1 % egg albumin solution + 2 ml plant extract at varying concentration and added 2.8 ml of PBS pH-6.4 was incubated at 37 o C for 15 mins and heated at 70 o C in water bath for 15 mins, let cool (RT) Absorbance at 660 nm.

Distilled water- Blank,

Control- everything without an extract

From the above experiment we are getting the following results:

1. Absorbance of Control is way less than the absorbance of Sample why?

2. Turbidity is increasing while increasing as the concentration of Plant extract how?

3. The paper shows distilled water as blank and some says PBS as blank. Why?

So kindly help me to conduct an anti-inflammatory activity of unknown crude extracts for egg albumin denaturation inhibition assay.