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C7661 Sigma

Collagen from rat tail

Bornstein and Traub Type I, powder, BioReagent, suitable for cell culture

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Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Components

All collagen molecules are composed of three polypeptide chains arranged in a triple helical conformation, with a primary structure that is mostly a repeating motif with glycine in every third position and proline or 4-hydroxyproline frequently preceding the glycine residue. Type I collagen differs from other collagens by its low lysine hydroxylation and low carbohydrate composition.

Other Notes

Collagen is classified into a number of structurally and genetically distinct types. We use the nomenclature proposed by Bornstein and Traub. Do not confuse Sigma type designations with recognized collagen classification types.

Packaging

10, 25, 50, 100 mg in poly bottle

5 mg in glass bottle

Preparation Note

This product was prepared by a modification of the extraction method of Bornstein, M.B., Lab. Invest., 7, 134 (1958). It is soluble in 0.1 M acetic acid at 1 mg/mL and should be stirred at room temperature for 1-3 hours until dissolved.

Application

Collagen from rat tail is used for the following applications:
• Immunohistochemistry
• Cellular activity assays
• Used in generation of dorsal root ganglion (DRG) explant cultures
• Used as one of the components during the preparation of the functionalized surface (in NMR setup)
• Used in cell culture (the glass coverslips were coated with nanowires at high concentrations mixed with collagen)
• Used for biofunctionalization of the microchannels

This product is intended to produce thin layer coatings on tissue culture plates to facilitate attachment of anchorage-dependent cells, recommended for use at 6-10 μg/cm2. It is NOT intended for production of 3-D gels. Type I collagen is often used in cell culture as an attachment substratum with myoblasts, spinal ganglia, hepatocytes, embryonic lung, heart explants, fibroblasts, endothelial cells, and islet cells have all been cultured successfully on films or gels of type I collagen. Collagen type I may also be used in research of Idiopathic pulmonary fibrosis (IPF), studies on the effect of ER stress IPF on lung fibroblasts. Collagen in acidic solution can produce three dimensional scaffolding with use in bioengineering and cell culture applications.

Biochem/physiol Actions

Collagen is an essential ingredient of connective tissue. Studies in a Chinese family show that mutation in COL1A1 (collagenase type I) is linked with type I osteogenesis imperfecta. Collagen is linked with subchondral turnover of bone, and might have potential as marker to determine the state of joint space narrowing and osteophytes in osteoarthritis.

General description

Col1a1 (collagen, type I, α1) or collagen is a major structural human protein, which assembles in the form of fibrils. It is a very long, thin and the most abundant protein found in the human body. It is a supercoiled right-helix of three left-handed polypeptide chains. These chains are composed of ~1040 amino acids, which are essentially repeats of three amino acids -(Gly-X-Y)n. Gly is glycine and X and Y can be any amino acids, but in humans are usually proline and hydroxyproline, respectively.

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Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport
WGK Germany 
1

Frequently Asked Questions

How can I sterilize collagen solutions?
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.
How are collagen solutions stored?
Collagen solutions should be stored in the refrigerator at 2-8 °C for up to 1 year, unless it becomes contaminated.
Why am I seeing some insolubles in my collagen solution?
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.
How can I make a 3-D collagen gel?
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 pH 6. 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.
How long can collagen coated plates be stored?
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.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
How do I find price and availability?
There are several ways to find pricing and availability for our products.  Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers. 
What is the Department of Transportation shipping information for this product?
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
  What kind of solution is the A5955 - Antibiotic Antimycotic Solution (100×), Stabilized product suspended in?
A5955 - Antibiotic Antimycotic Solution (100×), Stabilized is supsended in a sodium citrate buffer solution. Source: Israel production group
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Protocols & Articles

Articles

Cancer Stem Cells: New Targets for Cancer Therapy

Are stem cells involved in cancer? Cancer stem cells (CSC’s) are subpopulations of cancer cells that can self-renew, generate diverse cells in the tumor mass, and sustain tumorigenesis1. Cancer resea...
Keywords: Cancer, Cell attachment, Cell culture, Cell proliferation, Gastrointestinal, Gene expression, Transcription, transformation

Collagen Attachment Protocols, Solubility, and Stability

A1) 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...
George Sitterley
BioFiles 2008, 3.8, 5.
Keywords: Centrifugation, Dialysis, Filtration, Purification, Sterilizations

Collagen Usage Recommendations

Collagen is one of the most abundant proteins in the connective tissues and internal organs of mammals. It provides the tensile strength of the extracellular matrix (ECM). It is classified into a num...
George Sitterley
BioFiles 2008, 3.8, 4.
Keywords: Cell culture, Morphogenesis

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Peer-Reviewed Papers
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References

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