Q1. 

In the laboratory classes in weeks seven and

eight, you used western blotting to detect the MFN2 protein in a patient sample.

The expected western blot result is shown in Figure 1. Complete the steps below

(A-B) to upload and interpret your western blot results.

Figure 1.

Expected western blot result for detection

of MFN2 protein in patient sample. 1 = Precision Plus Kaleidoscope Prestained

Protein Standard, 2 = patient protein lysate.

A) RESULTS UPLOAD:

Upload a photograph of your

western blot result to the file upload box below.

B) RESULTS INTERPRETATION:

Compare your western blot

result to the expected result shown in Figure 1. Did your western blot produce

the expected result? (YES/NO) (4 marks)

If YES, proceed to the next question (Q2).

If NO, explain how your western blot result differs from the

expected result and suggest one or more sources of error or procedural issues

that could explain this difference. (2-3 sentences)

Q4. 

The MFN2 protein normally localises to mitochondria and is anchored in

the outer mitochondrial membrane (OMM), where it mediates mitochondrial fusion.

Would you expect the patient’s MFN2 protein to be present in the OMM? (1-2

sentences) (2 marks)

Q3.

  Which functional domains would you expect

to be present in the patient’s MFN2 protein? (select all that apply)

Q2.

Refer to your western blot result, or to the

result shown in Figure 1 if your western blot did not yield the expected result.

A)

Use the Precision Plus Kaleidoscope Prestained Protein

Standard to estimate the molecular weight of the patient’s MFN2 protein. (1

mark)

B)

Based on the patient’s DNA sequencing results, it

was predicted that the patient’s MFN2 protein would be truncated. Do the

western blot results support this prediction? Explain why or why not. (1-2

sentences) (2 marks)

Q1. 

In the laboratory classes in weeks seven and

eight, you used western blotting to detect the MFN2 protein in a patient sample.

The expected western blot result is shown in Figure 1. Complete the steps below

(A-B) to upload and interpret your western blot results.

Figure 1.

Expected western blot result for detection

of MFN2 protein in patient sample. 1 = Precision Plus Kaleidoscope Prestained

Protein Standard, 2 = patient protein lysate.

A) RESULTS UPLOAD:

Upload a photograph of your

western blot result to the file upload box below.

B) RESULTS INTERPRETATION:

Compare your western blot

result to the expected result shown in Figure 1. Did your western blot produce

the expected result? (YES/NO) (4 marks)

If YES, proceed to the next question (Q2).

If NO, explain how your western blot result differs from the

expected result and suggest one or more sources of error or procedural issues

that could explain this difference. (2-3 sentences)

How long will the PVDF membrane be incubated in the anti-MFN2 antibody working solution?

Use the dilution equation (C1V1 = C2V2) to calculate the volume (µL) of anti-MFN2 antibody that is needed to prepare 5mL of a 0.2µg/mL working solution.

Which type of detection method will be used in this laboratory to visualise MFN2 on the PVDF membrane?

You will use an anti-MFN2 antibody to probe the PVDF membrane and detect the MFN2 protein during western blotting. The anti-MFN2 antibody used in this laboratory is conjugated to:

Which of the following reagents are toxic in contact with the skin? (select all that apply)