Rat Collagen Type I

This product is prepared from rat tail tendons by modification of a published procedure. It is supplied as an aqueous solution in 20 mM acetic acid (~100 mg of protein per vial). Protein concentration was estimated by the biuret method. The vial contains approximately 100 mg of protein. Volume per vial: 22–25 mL.

The product ships on wet ice and storage at 2-8°C is recommended. The product may acquire a gel-like appearance. This is expected behaviour. Under such circumstances, it is possible to warm the product gently, in order to reconstitute the product fully, before use. Do not exceed 37°C, as higher temperatures may cause protein denaturation and loss of biological activity. Do not freeze. Optimal conditions for attachment must be determined for each cell line and application.

Note on solution behaviour: C3867 is supplied as an acidic solution in 20 mM acetic acid. In this form, the neat stock will not gel spontaneously at room temperature. However, once diluted into a neutral or near-neutral aqueous solution (e.g. sterile tissue culture grade water), the collagen will gel rapidly at room temperature (~20-25°C). All steps following dilution must therefore be performed on ice.

1. An appropriate volume of the collagen solution should be diluted to a working concentration of 0.01% using sterile, tissue culture grade water. Perform this dilution step on ice. Once diluted, the collagen solution will gel rapidly at room temperature - keep the diluted solution ice-cold at all times until immediately prior to application.

2. Coat dishes with 6-10 µg/cm². Allow the protein to bind for several hours at room temperature or 37°C, or overnight at 2–8°C. Note: Room temperature or 37°C incubation is appropriate at this surface coating stage only, after the diluted solution has been applied directly to the dish. Do not allow the diluted solution to sit at room temperature prior to application.

3. Remove excess fluid from the coated surface and allow it to dry overnight. If sterility has been compromised, the dried, coated surface can be sterilized easily by overnight exposure to UV light in a sterile tissue culture hood.

4. Rinse with sterile tissue culture grade water or a balanced salt solution before introducing cells and medium.

Yes, this product is suitable for use as a substrate for improved cell attachment. The material has not been specifically tested for attachment in fibroblast cultures. Please see the link below to review additional information available in the product datasheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/318/793/c3867dat.pdf

Rat collagen solution can be used as a coating for human cells, as indicated by various publications. The following publication in PubMed suggests that rat collagen can serve as a viable alternative to human collagen for various applications in cell culture and tissue engineering. https://pubmed.ncbi.nlm.nih.gov/18203067/

This product is prepared at a concentration of approximately 4 mg/mL. A package of 1 Vial will typically have a volume of approximately 35-40 mL. The exact concentration is lot specific and reported in the product Certificate of Analysis. Please see the link below to review a sample or lot specific Certificate:
https://www.sigmaaldrich.com/product/sigma/c3867#product-documentation

This collagen is intended for research use only, and is not approved for IVD purposes.

The recommended surface coverage for this product is 6?10 μg/cm2. Collagen solutions may be diluted to a working concentration of 0.01%. A general collagen coating protocol is available at https://www.sigmaaldrich.com/technical-documents/technical-article/cell-culture-and-cell-culture-analysis/mammalian-cell-culture/collagen-product-protocols

The collagen concentration of this product is reported on a lot specific basis and will be required to prepare the working solution. See the link below to review a sample Certificate of analysis:
https://www.sigmaaldrich.com/certificates/sapfs/PROD/sap/certificate_pdfs/COFA/Q14/C3867-1VL0000255665.pdf

Collagen solutions should be stored in the refrigerator at 2-8 °C for up to 1 year, unless it becomes contaminated.

The storage conditions that a Sigma-Aldrich catalog and label recommend for products are deliberately conservative. For many products, long-term storage at low temperatures will increase the time during which they are expected to remain in specification and therefore are labeled accordingly. Where short-term storage, shipping time frame, or exposure to conditions other than those recommended for long-term storage will not affect product quality, Sigma-Aldrich will ship at ambient temperature. The products sensitive to short-term exposure to conditions other than their recommended long-term storage are shipped on wet or dry ice. Ambient temperature shipping helps to control shipping costs for our customers. At any time, our customers can request wet- or dry-ice shipment, but the special handling is at customer expense if our product history indicates that the product is stable for regular shipment.

The solubility specifications for this product is clear to hazy colorless solution with a few insolubles at 1 mg/ml in water with 2 μl acetic acid (or 0.1 N acetic acid). The insolubles can be removed by settling or centrifugation.

Not all lots of C7661 are suitable for 3D gels. Product No. C4243 has been use-tested in this application. If C4243 is not available, we recommend that customers screen lots of C7661 to try to make 3D gels. We have found that collagen that has been sterilized by gamma-irradiation or by chloroform cannot be used to make 3D collagen gels. The following protocol can be used to make collagen gels: Materials required:Collagen gel solution 1.5 - 3 mg/mL 10X tissue culture medium containing phenol red Sodium bicarbonate or HEPES buffer Procedure: 1.Measure out 800 μL of collagen solution. 2.Add 100 μL of 10X medium (buffered with 1X sodium bicarbonate or HEPES) 3.Adjust pH with 1 N sodium hydroxide, if required (100 μl or less) 4.Add 10X medium to bring volume to 1 ml 5.Mix contents well. Solution should maintain red color to indicate physiological pH6. Dispense into wells to a depth of 1-2 mm (approx. 15 μL/well) 7.Transfer to 37 °C for 20-40 minutes 8.Examine for gel formation The above volumes represent quantities for use in 24 well plates. Volumes can be adjusted to accommodate any culture vessel.