Product Introduction
Collagen type Ⅰ from pigskin
(Cat# LV-Collagen004)
(For research only)
I Introduction
Collagen is the main component of pig skin, which has unique biochemical properties and physiological functions, and has good cell adaptability and cell proliferation. It can also promote wound healing, bone formation, and enhance skin metabolism. It can be used in a wide range of medical, cosmetic and cell culture applications. Our products are extracted and isolated from pig skin, purified in a 10,000-level laboratory through multiple processes, and maintain the biological activity of collagen. After testing, this product can be gelatinized after pH is adjusted to neutral, and can be used for cell culture of a variety of primary cells, including hepatocytes, vascular endothelial cells, mesenchymal stem cells, pancreatic beta cells, etc.
II Reagents and Materials
-Collagen mother liquor (this product Cat: LV-collagen004)
-Ultrapure ionized water
- Glacial acetic acid (Aladdin, CAS: 64-19-7; Cat:A116174-500ml)
-Sodium hydroxide (Sigma; Cat: S5881-500G)
-0.22μm membrane
-50mL disposable syringe
-Pipette and pipet tip
-Culture plate (dish)
- Biosafety cabinet
III Steps of Package
1. Add 172μl of glacial acetic acid to 100 mL of deionized water. In a biosafety cabinet, filter and sterilize with a 0.22μm membrane.
2. Add collagen mother solution of type I pigskin, so that the final concentration can be 50μg/ml. That is, add 50mg of collagen to hydrochloric acid solution per 1L (the corresponding volume of the collagen mother liquor is V=50mg/10mg/ml=5ml. Store at 4 °C for use.
3. Add collagen solution to the culture plate or Petri dish to be coated with an amount greater than/equal to 5μg/cm². Typically, the collagen solution added to the plate or dish is 60% of the recommended volume of medium. For example, the medium for 12-well plates is recommended to use a volume of 1 ml. Add 0.6 ml of collagen solution can be added to achieve saturation.
4. Incubate at room temperature for 1h in a biosafety cabinet.
5. Aspirate the collagen working solution. Place it in a biosafety cabinet to dry naturally and seal the parafilm. Store at 4 °C for no more than 3 months.
6. Before use, collagen-coated plates or Petri dishes need to be washed once with PBS or medium to remove residual acetic acid.
IV Gelatinization steps
1. Prepare 1M NaOH, 10×PBS or 10×DMEM, and the sterilized H2O, treat it sterile, and then pre-cool it on ice.
2. Preparation of gelatinous solutions:
① The added amount of collagen is:
② The added amount of NaOH:
Added amount of collagen×0.013μl=1M NaOH (μl)
③ The added amount of sterilized water:
final volume-volume of added collagen-volume of added NaOH-10×PBS or 10×DMEM = added amount of sterilized water3. Mix 10×PBS or 10×DMEM and NaOH solutions, sterilized water evenly on ice.
4. Add the collagen to the mixture to mix well and use immediately.
5. You can directly plate the mixed collagen solution when you use it. Incubate
at 37°C for 30 minutes and allow the gel to solidify before use.
Special Instructions
1. This product is a sterile liquid and no filtration is required for this product. Please feel free to use it.2. Collagen working fluid can be reused for 2-3 times, but possible contamination caused by multiple coatings should be avoided. If contamination is suspected, please stop using immediately.
3. We only detected 1mg/ml at the lowest gelatinization concentration, if a lower gelatinization concentration is required, you can test whether it can be glued by yourself.
4. When forming a gel, you only need to adjust the final solution to neutral, and the added amount of NaOH may be incorrect, which can be adjusted with the amount of water.
5. The mixing order of each component solution can be interchangeable, but NaOH cannot be added to collagen solution. Only collagen solution can be added to NaOH solution to prevent the inability of NaOH to mix rapidly and to produce a local gel.
V Contact Details
Tel: 0755-28284050
Technical Support: 19902901483 (Dr. Zhou)
References
【1】Bio-functionalization of porcine-derived collagen matrices with platelet rich fibrin: Influence on angiogenesis in vitro and in vivo[J]. Clinical Oral Investigations: 1-12.