Virtual Labs

Western Blot Analysis of the Cell Lysate using Antibody

1. After detection, how are protein bands typically quantified in Western blotting??

a: UV absorption Explanation

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b: Chemiluminescence Explanation

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c: Densitometry measurements Explanation

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d: Mass spectrometry Explanation

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2. If the primary antibody is an unmodified mouse monoclonal antibody, then the secondary antibody must be

a: anti-mouse IgG secondary antibody raised in horse Explanation

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b: anti-chicken IgG secondary antibody raised in mouse Explanation

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c: anti-goat IgG secondary antibody raised in horse Explanation

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d: anti-horse IgG secondary antibody raised in mouse Explanation

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3. Which of the following is NOT an application of Western Blotting?

a: Detection of post-translational modifications Explanation

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b: Protein quantification Explanation

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c: DNA sequencing Explanation

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d: Protein expression analysis Explanation

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4. The primary antibody in a Western blot is used to

a: Detect the secondary antibody Explanation

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b: Bind specifically to the target protein Explanation

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c: Transfer proteins from the gel to the membrane Explanation

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d: Visualize the separated proteins on the gel Explanation

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5. What is the purpose of the transfer buffer in Western blotting?

a: To separate proteins in the gel Explanation

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b: To facilitate the transfer of proteins from the gel to the membrane Explanation

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c: To block non-specific binding sites Explanation

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d: To enhance the antibody-antigen interaction Explanation

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6. Which washing buffer is commonly used in Western blotting to remove unbound antibodies?

a: TBS-T (Tris-buffered saline with Tween 20) Explanation

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b: PBS (Phosphate-buffered saline) Explanation

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c: SDS-PAGE running buffer Explanation

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d: Formaldehyde solution Explanation

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7. Which step is essential for ensuring proteins are transferred efficiently from the gel to the membrane in Western blotting?

a: Using a high-voltage power supply Explanation

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b: Properly equilibrating the gel and membrane in transfer buffer Explanation

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c: Staining the gel with Coomassie blue Explanation

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d: Incubating the membrane overnight at room temperature Explanation

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8. Why is it important to run a molecular weight marker alongside the samples in Western blotting?

a: To visualize the target protein Explanation

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b: To determine the size of the separated proteins Explanation

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c: To block non-specific binding Explanation

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d: To increase the sensitivity of the assay Explanation

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9. What is the primary purpose of the blotting step in Western blotting?

a: To separate proteins based on their size Explanation

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b: To transfer proteins from the gel to a membrane Explanation

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c: To denature proteins Explanation

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d: To detect specific DNA sequences Explanation

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10. What is the role of the secondary antibody in Western blotting?

a: To bind directly to the target protein Explanation

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b: To recognize and bind to the primary antibody Explanation

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c: To transfer proteins to the membrane Explanation

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d: To stain the proteins for visualization Explanation

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