Collagen I, Rat Tail
Cat.No.:IC-1921
Description
Collagen is a fibrous protein that consists of three α-chains which can combine to form a rope-like triple helix, providing tensile strength to the extracellular matrix (ECM) where it plays a key role in cell growth, differentiation, attachment, and cell migration. The α-chains containGXY repeats: glycine (G) is a small amino acid that fits well in the triple helix. X and Y are typically proline and hydroxyproline, which is critical for collagen stability. Type I is the most common fibrillar collagen (90%), and is mostly found in skin, bone, tendons, and other connective tissues. Rat tail collagen I can be prepared as a clear gel providing a 3D matrix or surface coated on tissue culture plates as a substrate for culturing primary cells such as keratinocytes and hepatocytes. Collagen I (Rat Tail) solution is supplied at 5mg⁄mL providing the flexibility to dilute to lower concentrations.
Use- Gelling Procedures
1. Place collagen (5mg/mL), sterile 10X phosphate buffered saline (PBS) or 10X Medium 199 (M199), sterile distilled water (dH2O), and sterile 1N NaOH on ice.
2. Determine the concentration and final volume of collagen needed for experimentation. See Example Calculation.
3. Determine the amount of reagents needed so that collagen is at the desired concentration in 1X PBS or M199 with normal osmolality and neutral pH. See Example Calculation.
4. In a sterile tube mix the dH2O, 1N NaOH, and 10X PBS.
5. Slowly pipet the collagen into the tube, and gently pipet solution up and down to mix well. The resulting mixture should achieve a pH of 6.5–7.5 (optimal pH is 7.0).
6. Dispense the collagen into the desired plates or dishes immediately and store them on ice. Gelling may occur rapidly at room temperature.
7. Incubate at 37°C in humidified incubator for 30–40 minutes or until a firm gel is formed.
8. Rinse the gel with sterile 1X PBS or cell culture medium before seeding cells.
Thin Coating Procedure
Note: Optimization for desired protein concentration may be required. A starting concentration of 5 μg/cm2 is recommended. Further dilution may be desired depending on the cell system.
1. Determine the volume needed for experimentation.
2. Dilute the collagen to 50 μg/mL in 20 mM acetic acid at the final volume needed.
3. Add solution to plates or dishes at 5 μg/cm2(e.g., 50 μg, or 1 mL of 50 μg/mL collagen is required for coating a 35-mm dish, which has a surface area of ≈10 cm2). Further dilution may be desirable for cell cultures requiring lower cellsurface adhesion strengths.
4. Incubate at room temperature for 1 hour.
5. Carefully aspirate solution from the well or dish.
6. Rinse dish three times with equal volumes of sterile 1X PBS or media to remove the acid.
7. Plates may be used immediately or air dried (stored at 2°C to 8°C) for future use.
Important Information
-Do not freeze.
-Perform manipulations on ice (2°C to 8°C) as gelling may occur rapidly at room temperature.
-We recommend that the following procedures be performed inan aseptic environment using aseptic techniques to preventcontamination.
Product Use
For Research Use Only. Not for use in diagnostic procedures.
Order Information
|
Cat./REF. |
Size |
Price($） |
Price(�) |
Price(￥/CNY） |
Price(￥/JYP） |
|
IC-1921-B |
10mg |
$120.00 |
� 144.00 |
￥1,200.00 |
￥23,400.00 |
|
IC-1921-C |
50mg |
$420.00 |
� 504.00 |
￥4,200.00 |
￥81,900.00 |
