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

1. Prepare a 2% (w/v) solution by dissolving gelatin in tissue-culture grade water.
2. Sterilize by autoclaving at 121 °C, 15 psi for 30 minutes.
3. Coat culture surface with 5-10 µL gelatin solution/cm² (i.e., 100-200 µg/cm²).
4. Allow to dry at least 2 hours before introducing cells and medium.
5. Alternatively, a gelatin solution (Product No. G1393) can be used to directly coat the cultureware.