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How to load equal amount of protein without using spectramax


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4 replies to this topic

#1 junbug

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Posted 18 October 2012 - 07:06 AM

Hi,

I was going to do BCA protein quantification with spectra max, but that machine is not working.
And I am not sure what I can do without it.
Is there any way?

Thank you so much for your help.

Edited by junbug, 18 October 2012 - 07:26 AM.


#2 Smog187

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Posted 18 October 2012 - 11:18 AM

You could still do the BCA protein quantification and run in parallel a known set of concentrations and then compare the color you get to your samples to get a "rough" estimation of protein concentration. Or you could run a SDS gel and load with known protein concentrations and compare your samples.

Edited by Smog187, 18 October 2012 - 11:18 AM.


#3 bob1

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Posted 18 October 2012 - 01:53 PM

You can also load based on cell number if you are using cell culture lysates, I find it more reliable than BCA.

#4 junbug

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Posted 19 October 2012 - 06:10 AM

You can also load based on cell number if you are using cell culture lysates, I find it more reliable than BCA.

I am using the mouse liver. So after homogenize and extract protein, I was planning to do the BCA, but that spectra machine is not working.Posted Image

#5 bob1

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Posted 19 October 2012 - 06:15 PM

If you have an ordinary spectrophotometer (even a nanodrop will work), you can do the assay using that, it is just a bit slower and you need to manually put the samples through rather than reading from a plate.




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