| Field | Specification |
|---|---|
| Mfr No | |
| Assay Time | |
| Detection Method | |
| Product Type | |
| Sample Type(s) | Biological samples (e.g. plasma, serum, cells) and cosmetic samples (e.g. collagen solution) |
| Shipping | |
| Species | |
| Storage |
Overview
For quantitatve determination of collagen in biological and cosmetic products. The assay uses FL375/465nm for signal readout. Compatible sample input includes Biological samples (e.g. plasma, serum, cells) and cosmetic samples (e.g. collagen solution). Typical stated assay timing is 80 min.
Key elements and design rationale
- Readout format: FL375/465nm supports plate-based signal acquisition and consistent comparison across matched samples.
- Sample compatibility: The stated sample scope includes Biological samples (e.g. plasma, serum, cells) and cosmetic samples (e.g. collagen solution), which is useful when aligning matrix type with calibration and control design.
- Analytical range context: The supplied specifications include a stated detection limit of 2 µg/mL for interpreting low-signal samples.
- Feature emphasis: Safe. Non-radioactive assay.
Additional feature notes highlight Fast and sensitive. Linear detection range (20µL sample): 2 µg/mL to 50µg/mLcollagen in 96-well plate assay; Convenient and high-throughput.”Add-mix-read” type assay. No wash and reagent transfer steps are involved. Can be readily automated for processing thousands of samples per day. Available format information for this listing includes 100 Tests.
Biological background
This product is centered on measurement of enzyfluo collagen within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.
More details
COLLAGEN is the key structural protein of connective tissue and the most abundant protein in mammals. It occurs in many different types and forms with Types I -V being the most common. Aside from the crucial role it plays in the body, it has numerous medical applications such as its use in reconstructive surgery including bone and skin grafts. It is also commonly used in cosmetics due to its anti-aging and skin healing properties. Assay methods available for quantifying collagen currently range from needing extensive hydrolysis procedures with acids and bases to using expensive antibodies and complicated protocols. BioAssay Systems collagen assay kit delivers a very simple, safe, and sensitive method to quantify collagen in samples. In the first step of this procedure, collagen in the sample is enzymatically digested into peptides. Subsequently, the N-terminal glycine containing peptides react with the dye reagent to form a fluorescent complex. The fluorescence intensity of this product, measured at λex/em = 375/465 nm, is directly proportional to collagen concentration in the sample.
Detection method
Fluorescent (FL 375/465 nm).
Detection limit and analytical sensitivity
Reported detection limit: 2 µg/mL.
Procedures and timing
Stated procedure or timing information: 80 min.
Research relevance and current trends
- Plate-based quantification and side-by-side group comparison remain central use cases for this assay format.
- The product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.
- Short assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.
Common research applications
- Quantify enzyfluo collagen in plasma, serum, cells by FL375/465 nm readout.
- Compare treatment or phenotype groups using matched plasma, serum, cells handling.
- Monitor time-course or pre/post changes in plasma, serum, cells across study conditions.
Interpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.
Notes for experimental interpretation
- Matrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.
- Use appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.
Does this assay measure insoluble collagen?
Yes, this assay can measure insoluble collagen. We would recommend using dried and ground powder sample (e.g. powdered bovine tendon). Below are the two methods that we have tested for insoluble collagen assay.
Method 1: Prepare sample at 1.0mg/mL or lower in 0.5 M NaOH. Incubate in a water bath at 37°C for 2 hours, shaking or vortexing to mix every 30 minutes. Neutralize with an equivalent volume of 0.5 M HCl then 10x dilute: 10 uL neutralized sample + 90 uL assay buffer. Use diluted solution for assay.
Method 2: Prepare sample at 1.0mg/mL or lower in 0.5 M NaOH. Sonicate sample 10 pulses, then heat at 100°C in water bath for 5 minutes. Sonicate another 10 pulses, and heat another 5 minutes at 100°C. Lastly again sonicate 10 pulses followed by neutralization with 0.5 M HCl then 10x dilute: 10 uL neutralized sample + 90 uL assay buffer. Use diluted solution for assay.
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. We have a hydroxyproline assay, DHYP-100.
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 laboratories requiring additional technical capacity, we provide scientific support services including assay execution, method guidance, product sourcing, and customization to align the assay with specific experimental objectives. If you need assistance selecting the appropriate kit configuration, adapting the workflow to your application, or identifying related research services, please click Talk to a Scientist, email support@biohippo.com, or review our Research Services; a member of our scientific team will follow up with recommendations tailored to your study.
Ginsenoside compound-Mc1 attenuates oxidative stress and apoptosis in cardiomyocytes through an AMP-activated protein kinase-dependent mechanism
Hong, S., et al. (2020). Ginsenoside compound-Mc1 attenuates oxidative stress and apoptosis in cardiomyocytes through an AMP-activated protein kinase-dependent mechanism.Journal of Ginseng Research.44(4): 664-671. Assay: Collagen in mouse heart tissue.
