C. Procedure for LDH Activity Assay The activity assays are set up and run in cu

Question

C. Procedure for LDH Activity Assay The activity assays are set up and run in cuvettes, so that you can measure the increase 10, 22.5 mM lactate, and 1 mM NAD, to which a small amount of the LDH sample will be added. The timing for starting and reading the assay must be exact, so read through in absorbance at 340 nm. Each cuvette will contain 90 mM CAPS buffer, pH the entire procedure before beginning. 1. Prepare a master mix by making a 1:10 dilution of the 10 mM NAD' into the 100 mM CAPS/25 mM lactate stock. You will need 1 ml of master mix for each assay, and you have four samples to run, plus a blank. Some of your samples will most likely need to be run more than once to get a good absorbance curve, so prepare enough master mix for 10 assays. Note that the master mix should be room temperature when you begin running the assays.

Explanation / Answer

Answer:

Preparation of master mix:

Master mix is required for 10 assays. 1ml for each assay x 10 = total 10ml required

So master mix sufficient for 10 assays (10ml) can be prepared by adding 1ml of 10mM NAD+ stock solution to to 9ml of 100mM CAPS/25mM lactate stock.

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