VitroCol®, Human Collagen, Solution, 3 mg/mL

Price(USD):$370.50

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Equl SKU Product Name UOM Specification Shipping Temperature Price(USD) Add to Cart:
BM-5007-20ML VitroCol®, Human Collagen, Solution, 3 mg/mL EA 20 mL Gel Ice $370.50  
Introduction:

VitroCol® collagen is the first widely available, naturally produced purified human collagen for research purposes. VitroCol® sets the standard for purity (>99.9% collagen content), functionality and represents the only native-like human collagen offered.

VitroCol® collagen is naturally secreted from human neo-natal fibroblast cells. The human fibroblasts are cultured in optimal conditions allowing the fibroblast to naturally and efficiently secret extracellular matrix. The extracellular matrix is then processed and purified to yield the naturally produced human collagen.

VitroCol® is approximately 97% Type I human collagen with the remainder being comprised of Type III collagen. It contains a high monomer content as measured by gel permeation chromatography.

VitroCol® is supplied at approximately 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. VitroCol® is soluble atelo-collagen in 0.01 N HCI, therefore, the pH is approximately 2.0.

VitroCol® is especially ideal for human cell culture systems when coating of surfaces, providing preparations of thin layers of culturing cells, or use as a solid gel. VitroCol® human collagen is provided in user-friendly packaging for use and storage. VitroCol® is sterile filtered and is supplied as a ready to use solution.

Parameter, Testing, and Method

VitroCol® Collagen Solution
Catalog #5007-20ML

Form

Solution

Package Size

20 mL

Sterilization Method

Filtration

Storage Temperature

2 to 10°C

Expiration Date

Listed on product label and
Certificate of Analysis

Concentration
(Biuret Protein Determination)

2.9-3.2 mg/ml

Purity
(collagen concentration)

> 99.9%

pH

Approx. pH 2, 0.01 N HCl

Gel Time
(Gel Time Assay)

< 90 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

Cell Attachment Assay

Pass

Source

Human, Neo-Natal, Fibroblasts

 

Preparation and Usage

Coating Procedure

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

  1. Transfer desired volume of collagen solution from the bottle to a dilution vessel as required. Dilute to desired concentration using sterile 0.01 N HCl solution. A typical working concentration may range from 10 to 100 ug/ml.
  2. Add appropriate amount of diluted VitroCol® material 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

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