re digests & re maps od and uses last class how to read a paper intro to paper 1
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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
+
+++
----
+
+++
----+
+++
----
+
+++
----
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
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