Collagen, Type I from Rat Tail is at a concentration of approximately 4 mg/mL in a 0.2M acetic acid solution (pH 3.0). Rat Tail collagen is soluble telo-collagen. This collagen product is provided in user-friendly packaging for use and storage. This product is sterile filtered and is supplied as a ready to use solution.
This product is ideal for coating of surfaces, providing preparation of thin layers for culturing cells, or use as a solid gel. Rat Tail Collagen is suitable for applications using a variety of cell lines including hepatocytes, fibroblasts and epithelial cells.
Parameter, Testing, and Method
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Rat Tail Collagen Solution
Catalog # 5056-20ML
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Form
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Solution
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Package Size
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20 mL
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Sterility
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No growth
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Storage Temperature
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2 to 10°C
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Expiration Date
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Listed on product label and Certificate of Analysis
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Concentration
(Biuret Protein Determination)
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3.8-4.2 mg/mL
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Purity
(collagen concentration)
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> 95%
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Polyacrylamide Gel Electrophoresis
(SDS – PAGE)
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> 85% collagen containing within alpha, beta,
and gamma bands < 15% collagen contained with bands traveling faster than alpha
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Endotoxin (LAL)
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< 1.0 EU/mL
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Cell Attachment Assay
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Pass
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Source
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Rat Tail Tendon
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Gelation Curve
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Characteristic
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Preparation and Usage
Coating Procedure
Note: Employ aseptic practices to maintain the sterility of the product throughout the preparation and handling of the collagen and other solutions.
- Transfer desired volume of collagen solution from the bottle to a dilution vessel if required. Further dilute to desired concentration using sterile 0.1% acetic acid solution. A typical working concentration may range from 10 to 100 ug/mL. Note: Use these recommendations as guidelines to determine the optimal coating conditions for your culture system.
- Add appropriate amount of diluted Rat Tail collagen to the culture surface.
- Incubate at room temperature or 37°C, covered, for 1-2 hours.
- After incubation, aspirate any remaining material.
- Rinse coated surfaces carefully with sterile medium or PBS, avoid scratching surfaces.
- Coated surfaces are ready for use. They may also be stored at 2-8°C damp or air dried if sterility is maintained.
3-D Gel Preparation Procedure
Note: It is recommended that the collagen and other working solutions be chilled and kept on ice during the preparation of the collagen.
- Place the following reagents on ice: – Type I Rat Tail collagen (4 mg/mL) – Sterile PBS (10X) – Sterile Cell Culture Water (dH2O) – Sterile 1 N NaOH
- Determine the final volume and concentration required of Rat Tail collagen required.
- Determine the amounts of reagents required to yield Rat Tail collagen at the concentration and pH required: a. Volume of Collagen needed = (Final Concentration) X (Total Volume) (Initial Concentration of Collagen)
- Volume of 10X PBS needed = Total Volume 10 c. Volume of 1N NaOH needed = Volume of Collagen X 0.017 mL
- Volume of dH2O needed = Total Volume – (Sum of a + b+ c)
- Mix the volumes calculated for 10X PBS, NaOH and dH2O in a sterile tube.
- Add Rat Tail collagen to the tube with the reagents and pipet up and down to mix. Vortexing is not recommended.
- Dispense the Rat Tail collagen mixture in the desired sterile plates or culture vessels.
- Incubate at 37°C for 1 hour for gel formation
Cell Attachment Bioassay
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