L7213-BC Collagen G

L7213-BC
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      Catalog Number AvailabilityPackaging Qty/Pack Price Quantity
      L7213
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          100 ml
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          Description
          OverviewGrowth and differentiation of cells in vitro are significantly influenced by the substrate used (glass or plastic). If culture flasks are coated with attachment factors such as Fibronectin, Collagen, Gelatine or Polylysine, better cell growth is often achieved. Collagen G is used in the production of gels, to embed cells. Collagen G gels are suitable for use as a substrate for adherent cells in a cell culture vessel and as a floating matrix in or on cell culture medium. 0.4% solution in 15 mmol/l HCl, type I, 4 mg/ml

          Collagen G for optimum geling
          We sell two different collagens: Collagen A (cat. no. L7220) and Collagen G (cat. no. L7213), acid-soluble calfskin collagens. Collagen G is used in the production of gels, to embed cells. Collagen G gels are suitable for use as a substrate for adherent cells in a cell culture vessel and as a floating matrix in or on cell culture medium. (Collagen A is used to coat culture flasks.) We have updated the recommendations for the manufacture of gels with Collagen G with regard to the concentration and the necessary pH values of the starting solutions, to optimise geling. New features include tips for examining gels under a microscope and taking pictures.

          Components for gel production
          Other components, also offered by us, are required for the production of Collagen G gels (cat. no. L7213). You can find an overview of these in Table 48.
          Tab. 48: Components required for gel production

          Recommendations for optimum geling with Collagen G

          Geling is heavily influenced by the pH value. Before starting work, pre-cool all reagents to a temperature of +2 - +8°C.

          Solution A:
          0.7 M sodium hydroxide and 1 M HEPES buffer (cat. no. L 1613) are mixed equally (e.g. 5 ml sodium hydroxide with 5 ml HEPES buffer).
          Solution B:
          A 10x medium and solution A are mixed equally (e.g. 5 ml medium with 5 ml solution A).
          • The pH of solution B should be between 7.90 and 8.05. It is advisable to check the pH using a suitable measuring method. If the gel should remain sterile, measure the pH of an aliquot previously removed.
          • 8.0 ml Collagen G is gently mixed with 2.0 ml of solution B to produce the ready-to-use solution for geling. This prevents the formation of air bubbles.
          • 3.0 ml of solution prepared in this way is pipetted into a 25 cm2 culture flask. For a culture flask with a diameter of 9 cm, 5.0 ml is required. The culture flask is then incubated vibration-free for at least 1 hour or, for optimum stability, 24 hours at +35°C (± 2°C).

          Examining the gel produced under a microscope and taking pictures
          If a 35 mm culture dish is filled with 2 ml Collagen G/buffer media mixture (pH approx. 7.2 - 7.8) and incubated, a gel with faint white diffusion is formed, viewed from above. This diffusion appears as a result of crosslinking of the collagen molecules and should not cause any problems when examining under a microscope or taking pictures.
          The accumulation of diffusion is proportional to the increase of thickness of the gel, for example if 5-7 ml instead of 2 ml per 10 cm2 Collagen G (35 mm culture vessel) is used. This could make microscopic and photographic analysis difficult.
          Specifications:

          Cell Attachment: Pass
          pH: 1.8-2.4
          Osmolality: 0.01-0.05 Osm/kg H20
          Collagen Content: 0.37-0.45%
          Hydroxyproline: 0.046-0.0565%
          Color: Colorless
          Appearance: Clear

          Catalogue NumberL7213-BC
          Brand Family Calbiochem®
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          Ship Code Ambient Temperature Only
          Storage +2°C to +25°C
          Do not freeze Ok to freeze
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