Really need help with this biochemistry problem.

[Raihan Mirza]

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Hi I am totally stumped in trying to solve this biochemistry problem. I would really appreciate it if someone would be able to help me.

When a biochemist is attempting to isolate or purify a single protein by chromatographic techniques, he/she often has to use a number of different columns with different properties in order to achieve the goal; i.e., a single column doesn’t do the job completely. Explain why, showing off your knowledge of different types of chromatography. In this case, let’s assume that the goal is to isolate a single protein of medium size (~12 kDa) with a net negative charge. How might you go about separating this protein from others that are larger/smaller, as well as those with positive or no charge?

Any help would be greatly appreciated.

 

[KeyzerSoze]

Ok... what are the different types of chromatography, and on what basis do they separate different proteins?

This really isn't rocket surgery.

 

[hoqhuuep]

Based on charge, use isoelectric focusing. Based on size, use SDS-PAGE. Doing both at the same time is called 2D gel electrophoresis... look it up on wikipedia or something.

 

[Hoju]

The possibilities are endless. If purification is the object then you could do a combo of gel filtration and anion exchange chromatography in different steps, some kind of affinity chromatography depending on the properties of the protein, or all or some of those in some sequence. Plus other options.....

 

[greatnt249]

1. Why is this thread in the allopathic forum?
2. How has asking for homework help not gotten this thread locked by now?

 

[Dral]

"Showing off your knowledge...".

lol

 

[MilkmanAl]

2. How has asking for homework help not gotten this thread locked by now?

Working on it.

 

[DrYoda]

Working on it.

Your mod powers must have a lag.

 

[MilkmanAl]

More importantly, they have forum constraints. 🙁

 

[engineeredout]

So the category of med school Biochemistry has been stretched to include college orgo?

 

[KeyzerSoze]

So the category of med school Biochemistry has been stretched to include college orgo?

Who said anything about med school biochemistry?

 

[fahimaz7]

Don't post that kind of crap in this forum. OMG. can someone help me with this electric flux? How about Friedel-Craft reactions? Anyone? Bueller?