I am working with C57BL/6 mice for the development of a T2DM mouse model. During the course of this experiment, the mice will have their urine, blood, and tissue samples collected and examined for the presence of specific biomarkers. These biomarkers will need to be accurately quantified. At present, I understand ELISA to be the gold standard for this, but are there any alternative assays that can be done?
I would suggest Bio-Plex Multiplex Immunoassays. Those assays can detect specific autoantibodies in a single run. If you have access to Bio-Rad Bio-Plex 2000 systems, you should be able to detect proteins, peptides and nucleic acid targets.
Hello Muhammed Zidan Khan
You may use Liquid Chromatography-Mass Spectrometry (LC-MS) where in you use liquid chromatography for the physical separation by segregating the molecules based on their chemical attributes such as size, polarity and affinity for the stationary phase, followed by identification and quantification of the separated molecules using their mass-to-charge ratios by mass spectrometry. LC-MS can target a wide range of biomarkers from metabolites to proteins and peptides. LC–MS is a bioanalytical method for quantitative analysis in biological fluids like serum, plasma, urine, etc.
For tissue homogenates, you may use Western Blot. However, the data produced with Western Blot is semi-quantitative. This is because it provides a relative comparison of protein levels, but not an absolute measure of quantity.
Best.
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