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Question regarding Quantity One for semiquantification of western

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Pippuri
United States

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Topic Started by Pippuri
on 7/14/2009 19:32 PM   
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I have to use this software, Quantity One (Bio-Rad), to quantify my western results... After going through all the steps and click on "report", I am very confuse with the numbers that I got--

I use local background substraction and all the bands are selected using boxes with equal size.It seems to me that "Adj. Vol. INT*mm2" is what I need for plotting, but I don't understand why the numbers are in negative values.

Any input from those who are familiar with this software are truly appreciated!


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Satinder
India

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Posted By Satinder
on 7/14/2009 23:44 PM   
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I have used Quantity one software. but never faced this type of problem.

Have you used the subtraction as per lane or whole gel?

Secondly, try using Filter Tab and transform Image to refine the image background. Perhaps that will help you out.

Satinder



heehawmcduff
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Posted By heehawmcduff
on 7/15/2009 10:09 AM   
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I've never used Quantity One, and you probably already know about this, but I've found ImageJ to be a great piece of software for analysing Westerns etc.  You can download add-ons which make the process very easy.  Just an alternative if your Quantity One problem remains...

http://rsbweb.nih.gov/ij/



Isadora
France

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Posted By Isadora
on 1/30/2012 4:41 AM   
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Hi Pippuri,
Have you solved your problem? Because i have the same.....
Thank for your answer



Pippuri
United States

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Posted By Pippuri
on 1/31/2012 8:58 AM   
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Hi Isadora,

I am still baffled by the negative value, but one thing I "sort of" figure out is to acquire the image under the UV light instead of the under the white light mode- that seems to somehow reduce the chance of getting negative value.... 

For the situation in which I couldn't get rid of the negative value, I realized that ignoring the negative sign and looking the numbers gave me reasonable correlation too...

I am not sure this is helpful, but please let me know how things turn out in yout hands.

Good luck



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