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Collagen I, rat tail

 
ALX-522-435-0020 20 mg 65.00 USD
 
ALX-522-435-0100 100 mg 150.00 USD
 
  • New formulation enhances gelling over a larger concentration range.

Product Specification

Alternative Name:COL1
 
MW:235kDa, 215kDa, 130kDa, 115kDa
 
Source:Isolated from rat tail tendons.
 
Concentration:5mg/ml
 
Formulation:Sterile liquid. In 0.02N acetic acid.
 
Purity:≥90% (SDS-PAGE)
 
Endotoxin Content:<100 EU/mg purified protein (LAL test)
 
Appearance:Hazy viscous liquid.
 
Activity: Tested in cell proliferation assay – Increased attachment of cells on collagen coated coverslips.
 
Application:Preparation of collagen gels, thin layer coating of surfaces (culture plates, slides, coverslips).
 
Short Term Storage:+4°C
 
Long Term Storage:+4°C
 
Handling:Keep sterile. Do not freeze.
 
Miscellaneous/General: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.
 
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
  1. Dilute 5mg/ml collagen to the desired concentration using sterile conditions. (Note: Lower concentrations will have reduced rigidity).
  2. Add enough collagen to cover the surface.
  3. Saturate a cotton ball or piece of filter paper with concentrated ammonium hydroxide and make a closed vapor chamber.
  4. Heat to 37°C until gel forms.
  5. Remove the ammonium hydroxide.
  6. Soak the gel surface in PBS for 1 hour.
  7. Wash several times PBS.
  8. Store the gel in PBS at +4°C until use.
  9. Equilibrate the gel in the desired media for 30 minutes prior to use.

NaOH Method (For 10ml)
  1. Sterilely add 1- 7ml of 5mg/ml collagen to a tube depending on desired final concentration. (Note: Lower concentrations will have reduced rigidity).
  2. Add 1ml of sterile 10X PBS.
  3. Add 1 ml of sterile 1N NaOH.
  4. Bring final volume to 10ml with sterile dH2O
  5. Cover the desired surface with a layer of the collagen and place at 37°C until the gel has solidified. Gel will only form id the pH is ≥7.0.
  6. Store in PBS at +4°C until use.
  7. Equilibrate the gel in the desired media for 30 minutes prior to use.
 
ALX-522-435 CHO
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.
ALX-522-435 SDS-PAGE
Coomassie stained SDS-PAGE. Lane 1, Molecular weight marker. Lane 2, 1.0µg Collagen I, rat tail.
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ALX-522-435 CHO ALX-522-435 SDS-PAGE

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),

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