Salivary Proteins Procedure Day II

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University of Louisville *

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331

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Biology

Date

Apr 3, 2024

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docx

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Uploaded by MajorMoon8437

Procedure Day II 1. Collect at least 2 mL of saliva. 2. Place 1 mL of saliva in a tube labeled “Protein” and add 0.1 mL of 10X sample buffer. 3. load 20  L of your sample into one of the wells in a protein electrophoresis gel. 4. Place 1 mL of saliva in another tube labeled “Amylase” and make the following dilutions: 1/10 = 250  L saliva + 2.25 mL TBE buffer, 1/300 = 100  L 1/10 dilution + 2.9 mL TBE buffer 5. add 900  L of the 1/300 dilution to another tube and mix with 0.1 mL of 10X sample buffer. 6. load 10  L of your sample into one of the wells in another protein electrophoresis gel. 7. Both gels will be treated differently for you to be able to answer the follow-up questions. Make sure you understand what each gel treatment involves and why it is done. 1. Based on the “Protein” gel, how many different proteins can you identify in your saliva sample? Based on the results from well 6, one protein was identified in the saliva sample 2. Based on the “Amylase” gel, which band do you think contains the amylase protein? The white band seen in lane 7, the only band, contains the amylase protein. This is because the total protein gel also contained only one band, indicating the presence of only one protein.

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Salivary Proteins Procedure Day II

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