How to draw graphs for SDS PAGE results?

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craterblizz
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How to draw graphs for SDS PAGE results?

Post by craterblizz » Thu Feb 23, 2012 3:47 pm

Ive got one gel with molecular weight marker: Phosphorylase B, BSA, Ovalbumin, Carbonic anhydrase, Lactoglobulin and Aprotinin.
Then ive got 4 gels with same protein of 150 Kda giving two bands of 50 KDa and 25 KDa. is there anyway i can draw graphs from this data?

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JackBean
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Post by JackBean » Thu Feb 23, 2012 6:25 pm

and what kind of graph do you want to make?
How is it that 150 KDa protein gives 50 and 25 KDa?

The only I can think of is calibration to determine exact size of your proteins.
http://www.biolib.cz/en/main/

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EnricoPallazzo
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Re:

Post by EnricoPallazzo » Thu Feb 23, 2012 7:27 pm

Looking for a calibration curve?
On the X-axis plot the migration distance of each band (in mm)
On the Y-axis plot the log of the molecular masses of the proteins

Then you`ll get a (hopefully) linear slope from which you can calculate the exact masses of the unknown proteins: http://chemistry.ewu.edu/jcorkill/biochem/PAGESDS.gif

JackBean wrote:How is it that 150 KDa protein gives 50 and 25 KDa?


--> IgG :mrgreen:

craterblizz
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Re: How to draw graphs for SDS PAGE results?

Post by craterblizz » Fri Feb 24, 2012 2:53 am

yes that is IgG (150 KDa) split into light and heavy chain fragment, hence 50 KDa and 25 KDa.

By migration distance you mean Rf value? dist travelled by the band/ distance travelled by the dye right? If yes, then the X axis value shouldn't be in 'mm' right?
Also, is there anyway i can find out the standard Rf values for the molecular markers i mentioned?

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canalon
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Post by canalon » Fri Feb 24, 2012 2:59 am

If your protein ladder was bought from any reputable company, it is likely to be available on their website. Just look the item number on their catalogue. Or give them a phone call.
Patrick

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