glass bottle of 5 mg
cell culture | mammalian: suitable
0.1 M acetic acid: 1 mg/mL (Allow to stir at room temperature 1-3 hours until dissolved.)
rat ... Col1a1(29393)
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We recommend transferring the collagen solution to a glass bottle with a screw cap and carefully layering chloroform at the bottom. The amount of chloroform to use should be ~10% of the volume of collagen solution. DO NOT SHAKE OR STIR. Allow to stand overnight at 2-8 °C. Aseptically remove the top layer containing the collagen solution to a new bottle. We do not recommend sterilizing the collagen solution by membrane filtration. We have found substantial protein loss by this method.
Collagen solutions should be stored in the refrigerator at 2-8 °C for up to 1 year, unless it becomes contaminated.
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.
We make no recommendations about storage of plates after coating. Since several factors such as humidity may impact longevity of coated plates, customers should make their own determination as to whether plates can be stored and under which conditions. We offer Sigma Screen collagen coated plates (Product No. S3315) that indicates the following: For optimal performance, the unopened product should be stored in a dry place at 2-8 °C. The product may be stored at room temperature for up to three months. The product should not be exposed to temperatures above 50 °C.
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
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Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
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