In lab: Notebook: 1. Titles of experiments and references to prot...

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In lab:
Notebook:
1. Titles of experiments and references to protocols in lab manual
2. Record the volume and buffer (composition) used for sonication and the sonicator settings used.
3. Record the volume of your crude extract
4 List the volumes and buffers (compositions) used for nickel chromatography
5. List the details of all ADH assays (note cuvet contents for blank and reactions, dilutions, dilution calculations, dA/min readings).

Post-lab:
1. Why is there a buffer difference between the purification steps and the enzymatic activity assays?
2. Calculate the enzyme concentration (in units/ml) and total enzyme units for your crude extract. Show your calculations.
3. In this lab, you were required to dilute your enzyme sample until your results fell within a linear range 0.08 0.12 dA/min. What if you used your initial undiluted assay results to calculate your enzyme concentration and total enzyme units? How does this compare to the results you got from your diluted enzyme results?
5. In the next lab period, you will check your load flow through and Ni+ 1st Wash samples for ADH activity
a. What results do you expect if the purification is working properly?
b. What results might indicate there is a problem with the purification?

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These solutions may offer step-by-step problem-solving explanations or good writing examples that include modern styles of formatting and construction of bibliographies out of text citations and references. Students may use these solutions for personal skill-building and practice. Unethical use is strictly forbidden.

1. Buffer for purification is dictated by type of chromatography and matrix used. Buffer for purification and matrix must be compatible. In other hand, buffer for enzymatic assay should not interfere enzimatic activity in any way. Those two buffers may match but don't have to...

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