RE digests & RE maps

OD and uses

Last class

How to read a paper

Intro to Paper 1

Purifying proteins

Purification – based on what properties?

Purifying proteins - Solubility

[Salt]

Fract

ion

pro

tein

in p

reci

pit

ate

+

+++

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+

+++

----+

+++

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+

+++

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Ammonium sulfate precipitation

Getting rid of salt: Dialysis

Ammonium sulfate precipitation

0-40%40-60%60-80%

Bradford assay & enzyme assay (why)?

- Every fraction

Lab

Dialysis

First evidence of PA-GFP…

Increase in fluorescence of PA-GFP…

pH affects

Photostability

OK – what about in cells?

Can we study dynamics?

Lets do a real experiment – live trafficking

Following live trafficking of proteins

Lysosomal trafficking needs microtubules