I am conducting the preliminary in vitro anti-inflammatory study on my plant sample using the BSA denaturation assay. My reaction mixture is 250ul BSA (4% w/v in PBS), 750ul PBS (pH 6.4) and 500ul Sample. The concentrations of my sample are in the range of 100ug/ml, 200ug/ml, 300ug/ml, and 500ug/ml. The absorbance was read at 660 nm. However, the OD for my Control is lower than that of my Samples, and the OD gradually increased with the increase in concentration.
This resulted in negative values for my inhibition percentage.
The formula I used is %inhibition=(Abs Control - Abs Sample)/Abs Control *100
Can anyone suggest to me what I might be doing wrong, and how can I overcome the problem
I took BSA alone as control, do one thing take BSA reading before and after incubation this will tell you if the protein denatured (turbidity after incubation at 72°C will increase if the protein denatured). And is your compound colored?
- Badges
- Science topic
- Similar topics
- Biological Science
- Botany
- Pharmaceutical Botany
- Plant Extracts