Western blot quiz | Abcam

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Western blot quiz

Test your knowledge. Do you know your western blot?

Question 1. Which sample can serve as a positive control for antibody binding in a western blot experiment?

a. A lysate from a knockout cell line or tissue

b. A lysate from a cell line or tissue expressing your protein of interest

c. Housekeeping proteins such as beta-actin, GADPH, beta-tubulin

d. All the above

Question 2. What is the first step of each western blot protocol?

a. Membrane stripping

b. Protein transfer

c. Antibody staining

d. Electrophoresis

Question 3. Why should you use a loading control for your western blot?

a. To ensure that all lanes in your gel were evenly loaded with the sample

b. To check for the even transfer from the gel to the membrane

c. To be able to publish your results in a good journal

d. All the above

Question 4. Why do we need a protein transfer step in western blot?

a. Eliminates high background

b. Membranes provide a superior surface for antibody binding compared to gels

c. Increases abundance of your target protein

d. Allows multiplexing

Question 5. If your western blot has no or weak bands, which troubleshooting tip should you try?

a. Increase the amount of protein loaded and/or the concentrations of antibodies

b. Increase the time of antibody incubation

c. Use Ponceau stain to ensure that proteins were transferred to the membrane

d. All the above

Question 6. If your western blot has a high background or more bands than expected, which troubleshooting tip should you try?

a. Titrate the amount of antibody used

b. Decrease the blocking incubation period

c. Increase the quantity of protein loaded

d. Decrease the number of washing steps

Question 7. What is the main advantage of fluorescent detection in western blot compared to chemiluminescent?

a. Signal amplification

b. Multiplexing

c. Antibody specificity

d. Less species cross-reactivity

Question 8. What can be the reason for you to get the western blot band sizes different from what expected?

a. Post-translational modifications

b. Post-translational cleavage

c. Alternative splicing and isoforms

d. All the above

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