protocolls

.
16.2. protocols Gail Kirk

Aim: to examine the relationship between absorbance and concentration of an
Enzyme and to calculate its specific activity.

Method:

16.2.1 Protein Assay- Lowry Assay
Verification of beers law/preparation of a Calibration Graph

9 test tubes were set up as follows

Test tube Containing
1 20µg protein + 80µg water
2 30µg protein + 70µg water
3 40µg protein + 60µg water
4 50µg protein + 50µg water
5 60µg protein +40µg water
6 70µg protein + 30µg water
7 80µg protein + 20µg water
8 100µg protein
9 100µg water (reagent blank)

Test tube nine was used to calibrate the machine. 5ml of alkaline reagent was then added to each test tube. The test tubes where then mixed and left for 10 mins. 0.5ml of Folin ciocalteau reagent was added to each test tube and the test tubes where then left for a further 30 mins after wh...