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Western Blot Controls and Antibody Selection

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Western Blot Controls and Antibody Selection

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Questions and Answers

What is the recommended boiling time for samples before SDS-PAGE?

10 minutes (correct)

5 minutes

20 minutes

15 minutes

What is the optimal loading control antibody used in the procedure?

Anti-tubulin antibody [mAbcam 9484]

Anti-GAPDH antibody [mAbcam 9484] (correct)

Anti-actin antibody [mAbcam 9484]

Anti-Bcl-2 antibody [mAbcam 9484]

What type of buffer is recommended for blocking during the procedure?

5% milk (correct)

2% BSA

10% BSA

10% milk

What could be a consequence of poor washing during the procedure?

Incomplete transfer of protein Signup and view all the answers

What is a possible issue when there are air bubbles between the gel and membrane during transfer?

Incomplete contact between gel and membrane Signup and view all the answers

Which type of blocking buffer would be more suitable for a different antibody than the Anti-GAPDH used?

5% BSA Signup and view all the answers

What could happen if too much protein or antibody is used in the procedure?

Decreased signal intensity Signup and view all the answers

How can incomplete contact between the gel and membrane during transfer affect the results?

Increase background noise on the membrane Signup and view all the answers

What is the purpose of diluting both sample and antibody when too much is used?

Enhancing signal strength Signup and view all the answers

Why might different blocking buffers be optimal for different antibodies?

Influencing binding specificity between antibodies and proteins Signup and view all the answers