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Determination of protein size via electrophoresis [View Printable]
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sarbol
Group: Member
Posts: 8
Joined: Dec 20, 2004
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The method simply involves determining the mobility of the protein in a polyacrylamide gel of known concentration (known pore size) together with a simple formula that relates the relative mobility to the ratio of protein size to the pore size of the gel.
Those intersted please refer to the article in
Elcetrophoresis, 25 (2004) pp2907-2911 |
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| Posted Dec 26, 2004, 14:11 PM |
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SueC.
Group: Member
Posts: 8
Joined: Jan 03, 2005
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Is this the same method as Ferguson plots that was developed by Rodbard and Chrambach back in the 70's? In their papers, they use increasing polyacryamide percentages. |
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| Posted May 04, 2005, 11:49 AM |
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sarbol
Group: Member
Posts: 8
Joined: Dec 20, 2004
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| SueC. said: | | Is this the same method as Ferguson plots that was developed by Rodbard and Chrambach back in the 70's? In their papers, they use increasing polyacryamide percentages. |
[answer=Sarbol] In some ways it is the same but uses a slit pore model for determination of the gel pore sizes (defined here as half width of the slit) using electrophoretic mobilty of proteins of known size. The equation used is very simple and relates the mobility to the size of the migrant molecule to that of the pore. Vice versa if a gel of known pore size (half width of slit) is used one can determine the size of the protien (whether globular or denatured). The values obtained for protein sizes are in close agreement with those found by more elaborate techniques like X-ray, light scattering, ultracentrifugation, GPC, etc. Besides one is able to monitor the variation of protein size during the course of its denaturation in a gel containing a gardient of urea concentration. |
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| Posted May 05, 2005, 0:30 AM |
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Long
Group: Member
Posts: 1
Joined: Sep 11, 2005
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I am going to use 2D gel to study protein but I don't know much about that. I want to get some documents about principles and protocol of 2D electrophoresis but I can't find them on internet. If you have any source of information please help. Thank you very much indeed! |
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| Posted Sep 12, 2005, 0:49 AM |
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sarbol
Group: Member
Posts: 8
Joined: Dec 20, 2004
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| Long said: | | I am going to use 2D gel to study protein but I don't know much about that. I want to get some documents about principles and protocol of 2D electrophoresis but I can't find them on internet. If you have any source of information please help. Thank you very much indeed! |
Dear Colleague: There are many books on 2D electroph. among which I mention a couple. 1. Gel electrophoresis of proteins_ A Practical Approach, Ed. B.D. Hames, IRL Press 1990 2. Proteomics from protein sequence to function- Chap. 3; Ed. S.R. Penington, BIOS Scientific Publishers LTD,2001 3. Proteomics in function and genomics-Protein structure analysis,pp 55-67, Birkhauser Verlag, 2001 4. Also there is a chapter on Urea gradient gels in the first book (Chap 1, pp 119-127) , in case you are interested. I hope these can be of help. |
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| Posted Sep 18, 2005, 4:46 AM |
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JKF
Group: Member
Posts: 1
Joined: Feb 02, 2006
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| SueC. said: | | Is this the same method as Ferguson plots that was developed by Rodbard and Chrambach back in the 70's? In their papers, they use increasing polyacryamide percentages. |
Actually the Ferguson of the Ferguson plot fame was my father, Kenneth A Ferguson; he developed the technique in the early 1960s when electrophoresing pituitary extracts to study growth hormone and published it in 1964. He is now 84 years and still going strong! |
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| Posted Feb 02, 2006, 7:54 AM |
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sarbol
Group: Member
Posts: 8
Joined: Dec 20, 2004
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Dear JKF:
I am glad to know you and even more happy to hear about your father. I wish him all the best and a very happy time. I take this opportunity to convey some words, if you won't mind.
One of our poets (Saadi of Shiraz- who was greatly loved by the father of Sadi Carnot the French scientist whose name we see in thermodynamics thus the name of the son) says:
"At the start the good was small, everyone after added slightly to it and it thus grew to be so enormous we now observe".
Another word from Ali (who is revered by the Iranians as the greatest human in the history) says: "There is only one vessel that not only never gets full rather it expands as more is placed in it, and the vessel is Science"
I don't quite know if I have translated these words properly but I hope you can get the great messages they convey.
The kindling your father lighted up has gorwn and will grow and illluminate further and further in time.
Science does not recognize boundries it is a universal treasury belonging to all across the globe.
So I am glad to have been able to continue your father's work and contribute even to the slightest.
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| Posted Feb 07, 2006, 3:27 AM |
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