At first I treated interferon beta protein to the tumor bearing mouse then I collected the tumor and blood from the mouse. I already made the tumor sample with PBS and collected serum from blood. Now I want to check interferon beta protein concentration in mouse tumor and serum sample by ELISA.
So, should I need to check the protein concentration by BCA assay at first? or only dilution with PBS and do the ELISA? example: I have 150 ul tumor and serum sample. If I want 10x dilution in one well (100 ul), can I just use 10 ul from sample and 90 ul PBS for one well?
Can anyone please suggest how can I make the sample for ELISA?
Dear Lata Rajbongshi,
the total protein levels are only required for normalization and can also be measured afterwards. The diution of your samples should be done in the same solvent that you use for the dilution of your interferon beta standards in the ELISA to provide a similar matrix backgtround. Make sure to measure the solvent as a blank as well.
Good luck - CL
Lata Rajbongshi : The following guide may help you
https://www.abcam.com/protocols/elisa-sample-preparation-guide-1
Dear Lata,
For tissue extraction, you can homogenize your samples in an extraction buffer, such as T-Per. You can use a tissue homogenizer for that. Then, centrifuge and collect the supernant; use this sample to measure in Elisa. Keep in mind you can further dilute your sample, in order to fit in the standard curve, but this dilution factor should be multiplied in the final Elisa result.
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