Does this assay measure hydroxyproline?
No, this assay does not measure hydroxyproline. This assay quantifies N-terminal glycine containing peptides from an enzyme based degradation of collagen.My standard curve isn’t turning out linear / I am getting inconsistent results?
You may also wish to incubate the digest mix step for an extra 30 minutes at 37°C (90 minutes total) to ensure that the digest reaction is complete.I’m not getting the same fluorescence readings for my standards as the standard curve on the protocol?
Fluorescence is measured differently depending on the plate reader. This is why plate readers use relative fluorescent units (RFUs) to quantitate the fluorescence readings. These are arbitrary units to assess the fluorescence intensity of the wells relative to each other. The numbers you see on the standard curve in the protocol are from the assay being performed on our lab’s plate readers. It is completely normal for your assay to produce different fluorescent readings; what is more important is to focus on the linearity of your standard curve.My sample, which I know has collagen, isn’t measuring any collagen with this assay?
Either your samples are diluted too much, or have not been diluted enough. We recommend running a serial dilution on your sample to determine the optimal dilution factor for quantification. This is the range at which dilutions start producing similar collagen concentration estimates after being corrected for dilution factor.Can I use this assay to measure cosmetic or beauty product collagen?
Yes, this assay can be used on store bought collagen. Because the collagen is often very concentrated, you will likely need to dilute it greatly (e.g.100-1000 fold) for it to be within the assay’s detection range.For more detailed product information and questions, please feel free to Contact Us. Or for more general information regarding our assays, please refer to our General Questions.