PorcineFlexiCol®, Solution, 3 mg/mL #5169-100ML

FlexiCol® is isolated from porcine hides and is a soluble atelo-collagen in 0.01 N HCI. FlexiCol® collagen is approximately 97% Type I collagen with the remainder being comprised of Type III collagen. FlexiCol® collagen is supplied at approximately a 3 mg/ml concentration. The concentration for each specific lot is provided on a Certificate of Analysis that is available with the purchase of each product. FlexiCol® collagen is ideal for coating of surfaces, providing preparation of thin layers for culturing cells, or use as a solid gel. FlexiCol® collagen is provided in a user-friendly packaging for use and storage. FlexiCol® is sterile filtered and is supplied as a ready to use solution.

Parameter, Testing, and Method	*FlexiCol®* Collagen Solution Catalog # 5169-100ML
Sterilization Method	Filtration
Form	Solution
Package Size	100 ml
Storage Temperature	2-10 °C
Expiration Date	Listed on product label and Certificate of Analysis
Concentration (Biuret Protein Determination)	2.9-3.3 mg/ml
pH	Approx. pH 2, 0.01 N HCl
Gel Time (Gel Time Assay)	< 40 minutes
Fibrillogenesis (Fibril Formation)	>0.5 Absorbance Units
Polyacrylamide Gel Electrophoresis (SDS – PAGE)	> 85% collagen containing within alpha, beta, and gamma bands < 15% collagen contained with bands traveling faster than alpha
Sterility	No growth
Endotoxin (LAL)	< 1.0 EU/ml
Osmolality	~ 30 mOsm
Cell Attachment Assay	Pass
Source	Porcine Hide – Pepsin Extracted

Coating Procedure

Note:Use these recommendations as guidelines to determine the optimal coating conditions for your culture system.

Remove required quantity of collagen from the bottle and dispense into a dilution vessel.
Dilute FlexiCol® in water to ~50 to 100 µg/ml (~1:30). A 0.01 N HCl solution may also be used.
Swirl contents gently until material is completely mixed.
Add appropriate amount of diluted FlexiCol® material to the culture surface ensuring that the entire surface is coated.
Incubate at room temperature, covered, for 1-2 hours. Aspirate any remaining material. Alternatively, incubate at room temperature until surface is dry.
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

Slowly add 1 part of chilled 10X PBS or 10X culture media to 8 parts of chilled collagen solution with gentle swirling.
Adjust pH of mixture to 7.2–7.6. Use of sterile 0.1 M NaOH is recommended. Monitor pH adjustment carefully (pH meter, phenol red, or pH paper).
Adjust final volume to a total of 10 parts with sterile water.
To prevent gelation, maintain temperature of mixture at 2-10°C.
To form gel, warm to 37°C. For best results allow approximately 90 to 120 minutes for gel formation.

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 FlexiCol®	With *FlexiCol®*

Human Umbilical Vein Endothelial Cells (HUVEC) 10X

VERO Epithelial Cells 10X

Human Dermal Fibroblast Cells 10X

MDCK Epithelial Cells 10X

This product is for R&D use only and is not intended for human or other uses. Please consult the Material Safety Data Sheet for information regarding hazards and safe handling practices.

FlexiCol®

FlexiCol® – Porcine Collagen Solution, Type I, 3 mg/ml, 100 ml

Catalog #5169-100ML (formerly #5169-B)

Directions for Use

Product Brochure

SDS

Certificate of Analysis

3D Gel Video

Full Product Description

Preparation and Usage

Cell Attachment Bioassay

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