Quantitative measurement of total protein in acid hydrolyzates of fresh, frozen, fixed and paraffin embedded tissues in a fast and simple 96-well plate format!
The QuickZyme total collagen and hydroxyproline assays are frequently used to determine the amount of collagen in biological samples such as tissues, tissue extracts, cell extracts and culture media. For the proper interpretation of the data obtained these should be compared to some reference quantity like wet- or dry- weight of the tissue, amount of protein, DNA etc. Some of these quantities are not easy to measure, or require an additional sample because they cannot be measured directly in acid hydrolyzates. It would be helpful if it was possible to determine such a reference in the acid hydrolyzate also used for determination of total collagen or hydroxyproline, such that no longer additional samples or cumbersome methods would be required. The QuickZyme Total Protein Assay offers a convenient solution in such cases.
In this assay the total amount of amino acids present in the same acid hydrolyzate used in the QuickZyme total collagen and hydroxyproline assays is easily determined. This total amount of amino acids in the hydrolyzate is a direct measure of the amount of protein present in the original sample. Combination of the QuickZyme total protein assay with either the total collagen or hydroxyproline assay allows the expression of data as collagen or hydroxyproline per total amount of protein.
The QuickZyme total protein assay is based on the formation of a blue colored product from genipin with free amino acids (Lee et al. Anal. Chim. Acta 480(2003)267; Fujikawa et al. J. Ferment. Technol. 65(1987)149). The assay measures the total amount of amino acids present in the hydrolyzed sample, with exception of proline and hydroxyproline. The amount of amino acids is a good measure for the amount of protein. A hydrolyzed protein standard is supplied with the kit to directly convert data to mg of protein.
The assay is simple and results in a colored product with an absorbance maximum close to 570 nm. The assay is developed to measure protein in acid tissue and/or protein hydrolyzates in such a way that the assay can be performed directly on the hydrolyzate without a drying or neutralization step. When used in addition to the QuickZyme Total Collagen, or QuickZyme Hydroxyproline assay kits, this assay kit allows for easy expression of the collagen or hydroxyproline amounts per total amount of protein.
- Quantitative measurement of protein in acid hydrolyzates
- Species independent
- Colorimetric read out at 570 nm
- Broad dynamic range: 0.05 to 3.0 mg/ml. Sensitivity: 0.007 mg/ml
- Uses BSA as standard
- Ease-of-use: faster and easier than ELISA
- Storage at room temperature
- This kit does not require special equipment
- Tissue extract, acid hydrolyzates of tissues, Conditioned media, Cells/cell extracts, Frozen or paraffin tissue sections
A typical protein standard curve in the range of 0.047-3.00 mg/ml protein, the background is not subtracted).
Unopened kit: Store at 4°C in the dark. Do not use kit components past kit expiration date.
Opened kit / reconstituted reagents: The reconstituted reagents should be stored light protected at 4°C (< 1 week), longtime storage at -20°C (stable for at least 2 months). The diluted color reagent solution should be prepared fresh and used the same day.
Application note 1: How to choose your collagen assay?
Application note 2: How to measure collagen in mouse tissues?
Application note 3: Collagen per protein analysis in fresh, frozen and formalin-fixed tissues
Application note 4: Collagen analysis in fish tissue
Ilex Life Sciences LLC is an official distributor of QuickZyme Biosciences.