I Introduction
Collagen type Ⅰ from rat tail is an important extracellular matrix that promotes cell adhesion and maintains cell differentiation [1,2]. Our products are extracted and separated from the tail tendon of SD rats and purified through multiple processes in a 10,000-level laboratory, which can maintain the biological activity of collagen. After testing, this product can be used for cell culture of a variety of primary cells, including hepatocytes, vascular endothelial cells, mesenchymal stem cells, islet beta cells, etc.
II Reagents and Materials
-Collagen mother liquor (this product Cat: LV-collagen001)
-Ultrapure ionized water
- Glacial acetic acid (Aladdin, CAS: 64-19-7; Cat：A116174-500ml）
-0.22 μm membrane
-50mL disposable syringe
-Pipette and pipet tip
-Culture plate (dish)
- Biosafety cabinet
III Steps of Package
1. Add 172 μl of glacial acetic acid to 100 mL of deionized water. In a biosafety cabinet, filter sterilize with a 0.22 μm membrane.
2. Add collagen mother solution of type I rat tail, so that the final concentration can be 50 μg / ml. That is, 5 mg of collagen per 100 ml of collagen working solution. Store at 4 °C for use.
3. Add collagen working solution to the culture plate or Petri dish to be coated with an amount greater than/equal to 5 μg/cm2. Typically, the collagen working 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. And 0.6 ml of collagen working 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 Special Instructions
1. This product is a sterile liquid and no filtration is required for this product. Please use it with confidence.
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.
References
【1】 Linsley, C., Wu, B. & Tawil, B. The effect of fibrinogen, collagen type I, and fibronectin on mesenchymal stem cell growth and differentiation into osteoblasts. Tissue engineering. Part A 19, 1416-1423, doi:10.1089/ten.TEA.2012.0523 (2013).
【2】 Sawada, N. et al. Effects of extracellular matrix components on the growth and differentiation of cultured rat hepatocytes. In vitro cellular & developmental biology : journal of the Tissue Culture Association 23, 267-273 (1987).
V Customer Service
If you find any quality problems with the product, please collect the original data and contact the company's salesmen or technical support at the first time. The company ensure after-sales service. Every laboratory has different conditions, different operating habits, different proficiency, and objective factors in experimental failure. If the operation is not strictly in accordance with the instructions or exceeds the time limit of after-sales, the company does not do after-sales. Please understand and support us.
Validity period and raw data provided:
Contamination problems: Within 96 hours of contamination, microscope photographs of differences should be provided.
IV Contact Details
Tel: 0755-28284050
Technical Support:19902901483 (Dr. Zhou)
