Collagen, Acid Soluble, Rat Tail, Collagen Solution, 4 mg/mL

Price(USD):$195.00

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BM-5056-20ML Collagen, Acid Soluble, Rat Tail, Collagen Solution, 4 mg/mL EA 20 mL Gel Ice $195.00  
Introduction:

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

Rat Tail Collagen Solution
Catalog # 5056-20ML

Form

Solution

Package Size

20 mL

Sterility

No growth

Storage Temperature

2 to 10°C

Expiration Date

Listed on product label and Certificate of Analysis

Concentration
(Biuret Protein Determination)

3.8-4.2 mg/mL

Purity
(collagen concentration)

> 95%

Polyacrylamide Gel Electrophoresis
(SDS – PAGE)

> 85% collagen containing within alpha, beta,
and gamma bands < 15% collagen contained with bands traveling faster than alpha

Endotoxin (LAL)

< 1.0 EU/mL

Cell Attachment Assay

Pass

Source

Rat Tail Tendon

Gelation Curve

Characteristic

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.

  1. 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.
  2. Add appropriate amount of diluted Rat Tail collagen to the culture surface.
  3. Incubate at room temperature or 37°C, covered, for 1-2 hours.
  4. After incubation, aspirate any remaining material.
  5. Rinse coated surfaces carefully with sterile medium or PBS, avoid scratching surfaces.
  6. 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.

  1. 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
  2.  Determine the final volume and concentration required of Rat Tail collagen required.
  3.  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)
  4. Volume of 10X PBS needed = Total Volume 10 c. Volume of 1N NaOH needed = Volume of Collagen X 0.017 mL
  5. Volume of dH2O needed = Total Volume – (Sum of a + b+ c)
  6.  Mix the volumes calculated for 10X PBS, NaOH and dH2O in a sterile tube.
  7.  Add Rat Tail collagen to the tube with the reagents and pipet up and down to mix. Vortexing is not recommended.
  8. Dispense the Rat Tail collagen mixture in the desired sterile plates or culture vessels.
  9. Incubate at 37°C for 1 hour for gel formation

Cell Attachment Bioassay

To demonstrate cell attachment, cells were seeded onto surfaces coated with the matrix product and onto negative control surfaces without the matrix product. Cells were seeded onto surfaces using serum-free DMEM media. All surfaces were blocked with a solution containing 1% BSA. Cells were then allowed to attach for one (1) hour at 37°C. The culture surfaces were washed with DPBS. The results indicate significant cell attachment with surfaces coated using the matrix product.
 
Without UltraPure® With UltraPure®
Human Umbilical Vein Endothelial Cells (HUVEC) 10X
VERO Epithelial Cells 10X
Human Dermal Fibroblast Cells 10X
MDCK Epithelial Cells 10X

 

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