Please provide a clear explanation. Thank you. I have an affinity purified sampl

Question

Please provide a clear explanation. Thank you.

I have an affinity purified sample of Arginase. I now run the affinity purified sample through a cation exchange column and collect the flowthrough and eluate. The SDS-PAGE gel for all three samples (affinity purified, anion exchanger eluate and anion exchanger flowthrough) shows a single band of 150 kD. I perform a Western blot with these, and measure the amount of urea produced by equal volumes of the three samples. My data is shown below. Based on all these data, what is your BEST conclusion (assume everything is referred to at pH 7)?

Explanation / Answer

- From the given information, the following points can be concluded. They are

-- The author tried to purify the enzyme arginase using three chromatograpgy systems i.e. anion exhange eluate, anion exchange flowthrough and affinity purified.

-- From the SDS-PAGE analysis it is observed that the enzyme Arginase is detected only through anion affinity and anion exchanger flowthrough as 150kD.

-- From the western blot annalysis of the sample the urea produced is more in the affinity processed sample. Arginase enzyme is primarily known for its role in the urea pathway. It is the final enzyme in the urea frmtaion. Hence the more urea produced the higher the presence of enzyme.

--Thus, affiniy purification system could more efficiently purify the enzyme arginase in comparison to the other two systems (Affinity purification system makes use of specific binding interactions between the molecules).

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