Nutragen®, Bovine Collagen, Solution, 6 mg/mL

Price(USD):$500.50

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BM-5010-50ML Nutragen®, Bovine Collagen, Solution, 6 mg/mL EA 50 mL Gel Ice $500.50  
Introduction:

Nutragen® collagen is known as the standard of all collagens for purity (>99.9% collagen content), functionality, and the most native-like collagen available.  Nutragen®  is isolated from bovine hides sourced from the only controlled, closed herd in the United States. Advanced BioMatrix’s manufacturing processes comply with stringent quality standards that have proven to yield unsurpassed lot-to-lot consistency.

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

Nutragen®
 collagen is supplied at approximately a 6 mg/mL concentration to provide an extremely firm gel.  The concentration for each specific lot is provided on a Certificate of Analysis that is available with the purchase of each product.  Nutragen® is soluble atelo-collagen in 0.01 N HCI, therefore, the pH is approximately 2.0.

Nutragen® collagen is ideal for coating of surfaces, providing preparation of thin layers for culturing cells, or use as a solid gel.  Nutragen®  collagen is provided in a 50 mL volume and is contained in user-friendly packaging for use and storage.  Nutragen®  is sterile filtered and is supplied as a ready to use solution.

Parameter, Testing, and Method

Nutragen® Collagen Solution
Catalog # 5010-50ML

Sterilization Method

Filtration

Form

Solution

Package Size

50 mL

Storage Temperature

2-10 °C

Expiration Date

Listed on product label and
Certificate of Analysis

Concentration
(Biuret Protein Determination)

5.8-6.8 mg/mL

Purity
(collagen concentration)

> 99.9%

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 bans < 15% collagen contained with bands traveling faster than alpha

Sterility

No growth

Endotoxin (LAL)

 < 1.0 EU/ml

Cell Attachment Assay

Pass

Source

Bovine Hide – Pepsin Extracted

 

 

Preparation and Usage

Coating Procedure

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

  1. Remove required quantity of collagen from the bottle and dispense into a dilution vessel.
  2. Dilute Nutragen®in water to ~50 to 100 µg/ml (~1:30). A 0.01 N HCl solution may also be used.
  3. Swirl contents gently until material is completely mixed.
  4. Add appropriate amount of diluted Nutragen® material to the culture surface ensuring that the entire surface is coated.
  5. Incubate at room temperature or 37°C, covered, for 1-2 hours.
  6. After incubation, aspirate any remaining material.
  7. Rinse coated surfaces carefully with sterile medium or PBS, avoid scratching surfaces.
  8. 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 or 10X culture media to 8 parts of chilled collagen solution with gentle swirling.
  2. 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).
  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. For best results 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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