Human Collagen, Type IV

Human Collagen, Type IV, Lyophilized, 5 mg

Catalog #5022

Type IV Collagen is the primary collagen found in the extracellular basement membranes separating a variety of epithelial and endothelial cells. It is a major component of the dermal-epidermal junction where it is mostly found in the lamina densa. It is a heterotrimeric molecule containing two α1-like and one α 2-like chains.

This Type IV Collagen is isolated from human placenta and is purified using a multi-step process. The product is supplied as a sterile, lyophilized powder containing 5 mg of Type IV collagen per vial. A Certificate of Analysis is available with the purchase of each product.

Type IV Collagen is typically used as a thin coating on tissue culture surfaces. Specific instructions are found in the Directions for Use. This product is generally used in vitro as a substrate scaffold to enhance cell attachment, adherence and proliferation. Type IV collagen may be used to culture epithelial, endothelial, muscle, nerve and many other cell types. Additionally, this product is suitable for use as a substrate for collagenase assays and positive controls.

Parameter, Testing, and Method	Type IV Collagen, Lyophilized Catalog # 5022
Form	Lyophilized Powder
Package Size	5 mg
Sterilization Method	Low Dose Irradiation
Storage Temperature	-20°C prior to reconstitution 2 to 10°C after reconstitution
Expiration Date	Listed on product label and Certificate of Analysis
Shelf Life	36 months
Shelf Life after Reconstitution	3 months
Polyacrylamide Gel Electrophoresis (SDS – PAGE)	Characteristic
Sterility	No growth
Cell Attachment Assay	Pass
Endotoxin (LAL)	< 1.0 EU/ml
Adventitious Agents	The human source material has been tested and found negative for hepatitis B virus, hepatitis C virus, and for human immunodeficiency virus-1 and -2 (HIV-1 and -HIV-2).
Source	Human Placenta

Preparation and Usage

Preparation Procedure

Reconstitute the 5 mg vial with 5 ml of cold, sterile 0.25% acetic acid and mix by pipetting up and down several times. Allow to sit at 2 to 8°C swirling periodically for > 1 hour.Note: The resulting solution will be slightly hazy. If there are some insoluble materials present and you wish to remove it, aseptically centrifuge the material.
Dilute the product to desired concentration with sterile 0.25% acetic acid. A typical final coating concentration may be 10 to 100 µg/ cm². Testing will likely be required to determine optimal concentrations required for different cell culture systems.
Add appropriate amount of diluted product to culture surface.
Incubate at room temperature or 37°C, covered, for 1-2 hours.
After incubation, aspirate any excess remaining material.
Rinse coated surface carefully with a sterile balanced salt solution. Avoid scratching surfaces.
Aspirate remaining material from coated surface.8. Coated culture vessels are now ready to use. The coated culture vessels may be stored at 2 to 10°C.

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 Human Collagen, Type IV	With Human Collagen, Type IV

Human Umbilical Vein Endothelial Cells (HUVEC) 10X

VERO Epithelial Cells 10X

Human Dermal Fibroblast Cells 10X

MDCK Epithelial Cells 10X

Disclaimer

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.

Human Collagen, Type IV