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1- please measure protein conc. for each samples
2- load equal protein (micro gram), let controls, transfer to blot and develop for protein X, scan it
3- detect band intensity using image J, i.e calculate mean band intensity for each sample. [ set the measurement to 8 bit, be sure that mean values are increasing with band intensity if there, otherwise invert image.
4- now again use your reference actin antibody to blot, scan and calculate mean value for all 5 sample
4- calculate avg of mean of protein X as you say / mean of reference protein for all 5 samples.
5- do this same for sick and resistance mouse samples.
6- now comparing these 3 values you would be able to see the change.
good luck
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In Topic: Using serum as the sample in Western blot
03 June 2012 - 01:43 AM
PhDMom, on 18 May 2012 - 09:16 AM, said:
Has anyone ever successfully used mouse serum as the sample in a Western blot? If so, what (if anything) did you use to dilute it?
Thanks!
Thanks!
Hi.. you can dilute it with water, PBS.
