Principals of Enzyme Catalysis
Hypothesis- If more time takes place after the start of a reaction, then less absorbency will take place in the spectrophotometer because more of the reaction has taken place
Materials-Reaction cocktail, Assay Solution, Diluted Catalase, Diluted Phosphate Buffer (all created using pre-lab preparations), permanent marker, 9 clean test tubes, (2) 1ml pipettes, (1) 5ml pipette, 1 liter water in several beakers, gloves, safety goggles, Spectrophotometer
Procedure-
1. First, make sure you have all the required materials, some of which will have to be made by following the pre-lab requirements in the lab, some of which needs to be done a week beforehand. 2. Make sure your gloves and safety goggles are on 3. label 6 test tubes (blank, 0, .5, 1, 1.5, 2) 4. Transfer 3ml of assay solution into each test tube 5. Transfer .3ml of dilute buffer to blank 6. Label one of the remaining test tubes "Con" and the other "Rxn" 7. transfer 1.8ml of reaction cocktail into each 8. put .3ml of dilute phosphate buffer to "Con" 9. remove .3ml from "Con" and add to tube labeled "0" 10. add .3ml of diluted catalase to "Rxn". Mix. start timer 11. remove .3ml from "Rxn" and add to ".5" when the timer reads 30 seconds. Mix 12. remove .3ml from "Rxn" and add to "1" when the timer reads 60 seconds. Mix 13. remove .3ml from "Rxn" and add to "1.5" when the timer reads 90 seconds. Mix 14. remove .3ml from "Rxn" and add to "2" when the timer reads 120 seconds. Mix
