Gelatin Coating Protocol for Cell Culture
Gelatin is a molecular derivative of collagen and is synthesized from the alkaline digestion of collagen from bovine skin. Gelatin is a hydrocolloid and is rich in glycine, proline and hydroxyproline, which imparts structural stability. Gelatin offers an ideal cell-attachment substrate for cell culture. The optimal concentration depends on cell type and research objective.
Important Notes before Gelatin Coating
- Ensure cells are healthy and in an adequate number.
- Gelatin solutions need to completely liquefy at 37 °C before coating.
- Gelatin solutions may be stored at 4 - 8 °C indefinitely; elevated temperatures cause hydrolysis and loss of integrity.
- The typical surface coverage concentration is 100 – 200 µg/cm2. Surface coverage concentration may differ with the cells being cultured. Optimal conditions for attachment must be determined for each cell line and application.
- Do not over-dry post coating as this disrupts the structure of gelatin and affects cell attachment.
Gelatin coating protocol for Cell culture
- Prepare a 2% (w/v) solution by dissolving gelatin in tissue-culture grade water.
- Sterilize by autoclaving at 121 °C, 15 psi for 30 minutes.
- Coat culture surface with 5-10 µL gelatin solution/cm2 (i.e., 100-200 µg/cm2).
- Allow to dry at least 2 hours before introducing cells and medium.
- Alternatively, a gelatin solution (Product No. G1393) can be used to directly coat the cultureware.
Product No | Description | Coating concentration | Dilute in |
---|---|---|---|
G1393 | Gelatin solution | 100‑200 μg/cm2 | Sterile water |
G9136 | Gelatin from porcine skin | 100‑200 μg/cm2 | Warm basal medium (without serum) |
G1890 | Gelatin from porcine skin | 100‑200 μg/cm2 | Warm water |
G7041 | Gelatin from cold water fish skin | 100‑200 μg/cm2 | |
G9391 | Gelatin from bovine skin | 100‑200 μg/cm2 |
Frequently Asked Questions for Coating Culture ware with gelatin
- What is the purity of gelatin products?
We do not determine the purity of fibronectin solutions. All our fibronectin powders are 70 - 90% pure.
- What is the best solvent to dilute gelatin powders?
The solvents that are compatible to dilute gelatins are provided in Table 1.
- How long can gelatin be stored?
Gelatin solutions can be stored indefinitely at 2 - 8 °C. Gelatin-coated plates can be stored at 2 - 8 °C for up to 4 weeks provided they are sealed well to prevent contamination and/or over-drying up of gelatin. Do not use the product if discoloration or cracks appear on the surface of the coated material.
- What is the significance of Bloom number for gelatins?
Bloom number was introduced by Oscar T. Bloom and indicates the stiffness of a gelatin film. It was first measured using a Bloom Gelometer by a controlled process to measure the rigidity of a gelatin film. A higher number indicates a stiffer product. The Bloom number/Bloom strength helps in choosing the right gelatin product for your applications.
Materials
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