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Protein electrophoresis

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satty

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Dear all,
I am asking few basic questions abt electrophoresis-paper using 4 amino acids-Alanine,glutamate,lysine,histidine
We did the experiment by first drawing a line in the center of the filter paper and putting the 4 aa's in the center line and drying them.Followed by soaking the teo ends of the paper in tris buffer(ours was ph 2) and we ran it in voltage 150 for 20 mins,followed by ninhydrin application and colour development
Questions are
Why wait for the solvent (buffer) to reach the centre line before starting the current?
(hint: why dry your amino acids into small spots with a hair dryer?)
How do you calculate the distance migrated,direction(+/-) and the net charge?

Regards,
Satty

.........................

Posted Apr 27, 2007, 12:32 PM
satty

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Please, can someone answer my question?

.........................

Posted Apr 28, 2007, 3:03 AM
satty

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No answer/s yet?

.........................

Posted Apr 28, 2007, 10:52 AM
Tony Rook

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Satty:

Here is a paper that may be of interest to you:

N. H. Martin and G. T. Franglen
The Use and Limitations of Filter-paper Electrophoresis
J Clin Pathol. 1954 May; 7(2): 87105.


And to answer your 2nd question about migration calculations please refer to this reference:

Stern JR, Mais RF, Boggs JD.
"U" Protein and Migration Ratios in Paper Electrophoresis. Clinical Chemistry. 1957 Vol 3, No 5, 599-608.

Additionally, here is a Pubmed Link to a list of references about Electrophoresis on Filter Paper. Most of the work on filter paper electrophoresis was done in the 1950's and 60's.

Hope this helps!

.........................
Tony Rook

Posted Apr 28, 2007, 18:21 PM
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