PRODUCT DESCRIPTION EPICULT®-B Type IV is found mainly in the , a layer of upon which the epithelium resides(1). All are characterized by a secondary structure consisting of three polypeptide chains wrapped around one Type IV Collagen, another in a triple helical arrangement(2). The different types of collagen vary in the primary amino acid sequence of their CellAdhere™ Human, polypeptide chains, with type IV collagen being comprised of two identical 1(IV) chains and one 2(IV) chain that spontaneously Lyophilized form the triple helix in solution(3). CellAdhere™ Type IV Collagen, Human, Lyophilized is isolated from human placenta and is purified using a multi-step process. Product is supplied as a sterile, lyophilized powder containing CATALOG #07002 5 mg 5 mg of Type IV collagen per vial. Endotoxin level is 0.1 EU/mL or less, as assayed by Limulus Amoebocyte Lysate assay. Donors have been tested and found negative for hepatitis B DIRECTIONS FOR USE surface antigen (HBsAg) and HIV-1 antibodies and/or HIV-1 Perform all steps employing aseptic techniques. antigen. However, this product should be considered potentially infectious and treated in accordance with universal handling Preparing Collagen Coated Tissue Cultureware precautions. 1. Reconstitute the entire CellAdhere™ Type IV Collagen, Human, Lyophilized vial (5 mg) with 5 mL of sterile 0.25% acetic acid to obtain a 1 mg/mL solution. RECOMMENDED FOR 2. Vortex until everything has been solubilized. This product is ideal for coating surfaces with a thin layer of protein 3. Dilute reconstituted Type IV Collagen solution with sterile to support cell attachment, but is not suitable for 3D cell culture 0.25% acetic acid to reach desired concentration. applications. This product is compatible with a wide variety of cell types, including, but not limited to, epithelial, endothelial, muscle Note: Different dilutions will need to be tested to determine the and nerve cells. The optimal protein concentration may vary optimal concentration required for each culture system. depending on the cell type being used, and therefore must be Typical coating concentrations range between titrated for best results. 10 - 100 µg/mL. 4. Add desired amount of diluted Type IV Collagen solution on the surface of the cultureware to be coated. STABILITY AND STORAGE For example, use 1 mL to coat a 35 mm tissue culture dish. CellAdhere™ Type IV Collagen, Human, Lyophilized 5 mg 5. Cover coated cultureware, to protect from contamination. Product stable at -20C for 6 months from date of receipt. 6. Incubate at room temperature (15 - 25C) for 1 - 2 hours. Once reconstituted in sterile 0.25% acetic acid, product stable at 7. Aspirate excess solution. 2 - 8 C for 3 months.  Avoid scratching the coated surface.

8. Rinse coated cultureware with sterile phosphate buffered ACCESSORY PRODUCTS saline (PBS). 9. Use coated cultureware immediately. Alternatively, keep PRODUCT CATALOG # sterile and store at 2 - 8C damp or air dried. Dulbecco’s Phosphate Buffered Saline (D-PBS), 37350 Without Ca++ and Mg++

REFERENCES

1. Bornstein, P and Sage, H. Structurally distinct collagen types. Ann Rev Biochem; 49: 957-1003, 1980. 2. Tanzer, ML. Cross-linking of collagen. Science; 180(86): 561-566, 1973.

3. Heino, J. The collagen family members as cell adhesion . BioEssays; 29: 1001-1010, 2007.

Please refer to the Material Safety Data Sheet for additional information.

FOR RESEARCH USE ONLY. NOT FOR THERAPEUTIC OR DIAGNOSTIC USE.

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