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New formulation enhances gelling over a larger concentration range.
Product Details
| Alternative Name: | COL1 |
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| MW: | 235kDa, 215kDa, 130kDa, 115kDa |
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| Source: | Isolated from rat tail tendons. |
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| Concentration: | 5mg/ml |
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| Formulation: | Sterile liquid. In 0.02N acetic acid. |
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| Purity: | ≥90% (SDS-PAGE) |
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| Endotoxin Content: | <100 EU/mg purified protein (LAL test) |
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| Appearance: | Hazy viscous liquid. |
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| Activity: | Tested in cell proliferation assay – Increased attachment of cells on collagen coated coverslips. |
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| Application Notes: | Can be used for preparation of collagen gels, thin layer coating of surfaces (culture plates, slides, coverslips). |
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| Shipping: | Blue Ice Not Frozen |
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| Short Term Storage: | +4°C |
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| Long Term Storage: | +4°C |
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| Handling: | Keep sterile. Do not freeze. |
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| Scientific Background: | Collagen is the main component in connective tissue and helps to provide support for tissues. It is made up of several classes, with Type 1 collagen being the most common. Type 1 collagen has a herterotrimeric triple helical structure made up of two alpha-1(I) and one alpha-2(I) chains that form into elongated fibrils which are extremely strong. These fibrils can be found in skin, tendons, ligaments, and other connective tissues. Type 1 collagen has been shown to be useful as a substrate that promotes cell growth and proliferation. Under acidic conditions the protein is soluble, however by raising the temperature and pH the solution forms into a solid gel that can be useful for cellular studies. It can also be dried to form a thin layer on solid surfaces such as plates, slides, or coverslips to aid in cell attachment. |
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| Protocol: | Firm Gelling Procedure
(Note: The ideal concentration for gelling is 1 - 5mg/ml. The end user will need to determine the appropriate concentration for their specific purpose).
Ammonium Hydroxide Gas Method
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Dilute 5mg/ml collagen to the desired concentration using sterile conditions. (Note: Lower concentrations will have reduced rigidity).
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Add enough collagen to cover the surface.
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Saturate a cotton ball or piece of filter paper with concentrated ammonium hydroxide and make a closed vapor chamber.
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Heat to 37°C until gel forms.
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Remove the ammonium hydroxide.
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Soak the gel surface in PBS for 1 hour.
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Wash several times PBS.
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Store the gel in PBS at +4°C until use.
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Equilibrate the gel in the desired media for 30 minutes prior to use.
NaOH Method (For 10ml)
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Sterilely add 1- 7ml of 5mg/ml collagen to a tube depending on desired final concentration. (Note: Lower concentrations will have reduced rigidity).
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Add 1ml of sterile 10X PBS.
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Add 1 ml of sterile 1N NaOH.
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Bring final volume to 10ml with sterile dH2O
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Cover the desired surface with a layer of the collagen and place at 37°C until the gel has solidified. Gel will only form if the pH is ≥7.0.
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Store in PBS at +4°C until use.
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Equilibrate the gel in the desired media for 30 minutes prior to use.
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| Regulatory Status: | RUO - Research Use Only |
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Glass coverslips were coated with or without collagen in 60% ethanol and allowed to dry overnight. CHO cells were plated at onto the coverslips and grown for two days at 37oC with 5% CO2. Cells were fixed in 10% buffered formalin. Coverslips were washed in PBS, mounted upside down onto microscope slides, and imaged with a 20X objective.
Coomassie stained SDS-PAGE. Lane 1, Molecular weight marker. Lane 2, 1.0µg Collagen I, rat tail.
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Induced pluripotent stem cell-derived smooth muscle cells increase angiogenesis and accelerate diabetic wound healing: J. Gorecka, et al.; Regen. Med.
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A filter-free blood-brain barrier model to quantitatively study transendothelial delivery of nanoparticles by fluorescence spectroscopy: E. De Jong, et al.; J. Control. Release
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General Literature References
Reconstituted rat-tail collagen used as substrate for tissue cultures on coverslips in maximow slides and roller tube: M.B. Bornstein, et al.; Lab. Invest. 134, (1958),
