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Enzyme Kinetics V - Experiment 16 Isolation Purification...

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: , , Experiment 16 Isolation Purification and Kinetics of . E coli Alkaline Phosphatase Part 5 Biochemistry 332L - - 3 5 09 - This week we will be running the SDS PAGE we prepared in the last ( ). lab or rather one much like it We will be using it to verify the purity of . your enzyme and confirm the molecular weight Materials Electrophoresis Buffer 4L 4X Sample Loading Buffer 25mL PAGE Gel 1 gel ° Water Bath set to 90 C Staining Solution / 50 10 Destaining Solution 50mL / 10 10 Destaining Solution 100mL / Gel Box Power Supply Procedure 1. Mix 30μL of your Stage IV enzyme sample with 10μL of 4X Sample Loading . . Buffer Repeat for each of the other stages you have Mix 15μL of the protein . standards with 5μL of 4X Sample Loading Buffer 2. . Unwrap the gel and place it in the gel box Remove the comb from the gel . and fill the box with Electrophoresis Buffer 3. ° . Heat the samples in the 90 C water bath for five minutes 4. . Inject 20μL of one of the samples into one of the wells on the gel Repeat for . the other samples 5. ,
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