Protein Analysis Techniques

The video tutorial by Chris explains the process of analyzing proteins in samples using gel electrophoresis. It covers the steps of separating proteins based on molecular weight, preparing samples, running the gel, transferring proteins to nitrocellulose paper, staining, blocking, and incubating with antibodies. The tutorial provides a comprehensive guide to protein analysis techniques.

What is the first step in analyzing proteins of interest in samples?

• Heating the samples

• Adding a running buffer

• Separating them on a gel based on molecular weight

• Staining with ponzu

Why is a standard with known molecular weight added to the gel lanes?

• To prevent protein degradation

• To maintain the flow of electricity

• To compare against the samples

• To increase the speed of separation

At what voltage are the samples run through the gel?

• 50 volts

• 70 volts

• 90 volts

• 110 volts

What is the purpose of transferring proteins to Nitro cellulose paper?

• To dry the proteins

• To change protein color

• To allow interaction with antibodies

• To increase protein size

What is used to ensure proteins have transferred to the paper?

• Secondary antibody

• Running buffer

• Primary antibody

• Ponzu stain

What is the purpose of blocking membranes in a specific buffer?

• To prevent non-specific binding

• To enhance protein size

• To dry the membranes

• To change protein color

At what temperature are the membranes kept overnight with the primary antibody?

• 90° C

• 37° C

• 4° C

• Room temperature

What is the secondary antibody made with?

• Ponzu stain

• Water

• 5% milk in tbst

• 10% milk in tbst

What is the final step in the protein analysis process described?

• Heating the samples

• Washing the membranes in tbst

• Staining with ponzu

• Running the gel