Signaling Pathway Assays
Collagen Assay Kit (BA0186)
- SKU:
- BA0186
- Product Type:
- Assay
- Instrument:
- Microplate Reader
- Sample Type:
- Biological Samples, Cosmetic Samples
- Research Area:
- High-Throughput Screening
Description
Collagen Assay Kit (BA0186) (BA0186)
The Collagen Assay Kit (SKU: BA0186) offers a simple, safe and sensitive fluorimetric method for the quantitation of collagen in samples. Collagen is the key structural protein of connective tissue and the most abundant protein in mammals, occurring in many types and forms, with numerous medical and cosmetic applications. In this procedure, collagen in the sample is first enzymatically digested into peptides, after which the N-terminal glycine-containing peptides react with a dye reagent to form a fluorescent complex. The fluorescence intensity of this product, measured at 375/465 nm, is directly proportional to the collagen concentration in the sample. The assay uses only 20 uL of sample, offers a linear detection range of 2-50 ug/mL and follows a convenient add-mix-read format with no wash or reagent-transfer steps, allowing it to be readily automated. It is intended for research use only.
| Product Name: | Collagen Assay Kit (BA0186) |
| SKU: | BA0186 |
| Detection Method: | Fluorimetric (lambda ex/em = 375/465 nm) |
| Detection Range: | 2 ug/mL to 50 ug/mL collagen in 96-well plate assay |
| Sample Type: | Biological samples and cosmetic products |
| Species Reactivity: | All |
| Assay Time: | Approximately 80 min (60 min digestion plus two 10 min incubations) |
| Kit Size: | 100 Assays |
| Equipment Required: | Microplate reader |
| Storage: | -20C |
| Shelf Life: | 6 months after receipt |
| Shipping: | Gel Pack |
A safe, sensitive fluorimetric assay for the quantitation of collagen. Collagen is first enzymatically digested into peptides, and the N-terminal glycine-containing peptides then react with a dye reagent to form a fluorescent complex measured at 375/465 nm, proportional to collagen concentration. The linear detection range is 2-50 ug/mL using a 20 uL sample.
- Safe, non-radioactive assay
- Sensitive and accurate; uses 20 uL sample with a linear range of 2 ug/mL to 50 ug/mL collagen
- Convenient add-mix-read format with no wash or reagent-transfer steps
- Readily automated to process thousands of samples per day
- Determination of collagen in biological samples
- Determination of collagen in cosmetic products
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Prepare a 50 ug/mL Collagen Standard by mixing 5 uL of the 3 mg/mL Collagen Standard with 295 uL dH2O, then prepare standards as shown in the table. |
| 2 | Transfer 20 uL Standards into separate wells of a black flat-bottom 96-well plate and 20 uL of each sample in duplicate into separate wells (one Sample and one Sample Blank). |
| 3 | Prepare Digest Mix for the Standards and Sample wells by mixing, per well: 35 uL Buffer and 0.5 uL Digest Enzyme. Add 30 uL Digest Mix to the Standards and Sample wells and 30 uL Buffer to the Sample Blank wells. Tap to mix, cover and incubate 60 minutes at 37C. |
| 4 | Add 40 uL Dye Reagent to all wells and incubate 10 minutes at 37C. |
| 5 | Add 8 uL Developer to all wells and incubate 10 minutes at 37C. |
| 6 | Read fluorescence at 375/465 nm. |
Subtract the blank value (#4) from the standard values and plot delta F against standard concentrations to determine the slope. [Collagen] = (F_S - F_B) / Slope x n (ug/mL), where F_S and F_B are the readings of the Sample and Sample Blank respectively and n is the dilution factor. If the sample fluorescence exceeds that of the 50 ug/mL standard, dilute in water and repeat, multiplying by the dilution factor. Conversions: 50 ug/mL equals 5 mg/dL or 50 ppm.
| Component | Quantity | Storage |
| Dye Reagent | 5 mL | -20C |
| Collagen Standard (3 mg/mL) | 40 uL | -20C |
| Buffer | 5 mL | -20C |
| Developer | 1 mL | -20C |
| Digest Enzyme | 70 uL | -20C |
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