my samples are whole blood samples. I separated the WBCs to detect the protein level of two heterogeneous nuclear ribonucleoproteins. Now I have the relative protein level but do not know how to interpret them. I used patient samples and healthy control samples and a housekeeping gene B actin.
the answers from here may be useful
How to analzye western blot bands? is there any full guide how to fully interpret the results? | ResearchGate
Hello,
after measuring the intensity of each band, you must normalise the values you get from your ribonucleoproteins to the values of the corresponding actin band. Last, you compare the normalised values of the ribonucleoproteins.
Here is a step by step guide how to do this.
https://licor.app.boxenterprise.net/s/kijrkoh8oqwv212577slsljldl74og16
Ideally in your case, I would use a total protein stain instead of a housekeeping protein. The expression of any housekeeping may differ from patient to patient or from patient to healthy individuals.
https://licor.app.boxenterprise.net/s/1q6nvqiov23om7n80hxo046w9mpfqx3s
There's really very little information in your question...
but unless you followed very specific conditions, a Western Blot using the standard setup is NOT quantitative at all - it's nice to look at as a confirmation and shows the protein band of the right size... but that's really it.
see e.g. Article Quantifying western blots: none more black
You'd at the very least needed to have loaded several dilutions of your samples and really should use a Chemiluminescence Imager to measure the output, best combined with fluorescently labelled antibodies instead of using a chemiluminescence reaction. Such a machine generally has software to do the quantification, so I assume you don't have access to one...
But why bother with all the work of a western blot when an ELISA is way quicker?
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