Different Title: COL1
MW: 235kDa, 215kDa, 130kDa, 115kDa
Supply: Remoted from rat tail tendons.
Focus: 5mg/ml
Formulation: Sterile liquid. In 0.02N acetic acid.
Purity: ≥90% (SDS-PAGE)
Endotoxin Content material: <100 EU/mg purified protein (LAL check) Look: Hazy viscous liquid. Exercise: Examined in cell proliferation assay – Elevated attachment of cells on collagen coated coverslips. Utility Notes: Can be utilized for preparation of collagen gels, skinny layer coating of surfaces (tradition plates, slides, coverslips). Delivery: Blue Ice Not Frozen Quick Time period Storage: +4°C Lengthy Time period Storage: +4°C Dealing with: Maintain sterile. Don't freeze. Scientific Background: Collagen is the primary part in connective tissue and helps to supply help for tissues. It's made up of a number of courses, with Kind 1 collagen being the most typical. Kind 1 collagen has a herterotrimeric triple helical construction made up of two alpha-1(I) and one alpha-2(I) chains that type into elongated fibrils that are extraordinarily robust. These fibrils could be present in pores and skin, tendons, ligaments, and different connective tissues. Kind 1 collagen has been proven to be helpful as a substrate that promotes cell progress and proliferation. Below acidic situations the protein is soluble, nevertheless by elevating the temperature and pH the answer types right into a strong gel that may be helpful for mobile research. It will also be dried to type a skinny layer on strong surfaces equivalent to plates, slides, or coverslips to assist in cell attachment.
Protocol: Agency Gelling Process (Observe: The best focus for gelling is 1 - 5mg/ml. The tip person might want to decide the suitable focus for his or her particular function). Ammonium Hydroxide Fuel Technique Dilute 5mg/ml collagen to the specified focus utilizing sterile situations. (Observe: Decrease concentrations may have decreased rigidity). Add sufficient collagen to cowl the floor. Saturate a cotton ball or piece of filter paper with concentrated ammonium hydroxide and make a closed vapor chamber. Warmth to 37°C till gel types. Take away the ammonium hydroxide. Soak the gel floor in PBS for 1 hour. Wash a number of instances PBS. Retailer the gel in PBS at +4°C till use. Equilibrate the gel within the desired media for half-hour prior to make use of. NaOH Technique (For 10ml) Sterilely add 1- 7ml of 5mg/ml collagen to a tube relying on desired closing focus. (Observe: Decrease concentrations may have decreased rigidity). Add 1ml of sterile 10X PBS. Add 1 ml of sterile 1N NaOH. Convey closing quantity to 10ml with sterile dH 2 O Cowl the specified floor with a layer of the collagen and place at 37°C till the gel has solidified. Gel will solely type if the pH is ≥7.0. Retailer in PBS at +4°C till use. Equilibrate the gel within the desired media for half-hour prior to make use of.