L7220-BC Collagen A, 1 mg/ml L7220

L7220-BC
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      L7220
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          1000 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. Culture flasks can either be coated with monomeric Collagen A or with fibrillar Collagen A; monomeric Collagen A is invisible thanks to its smooth surface and is thus more suitable for photographing. In each case, what form the Collagen A is in depends on the particular pH, the ion concentration and the temperature of the solution. We recommend different procedures depending on whether you wish to use the culture flask immediately or whether you provide a longer period of storage. 0.1% Collagen solution in HCl, type I, 1 mg/ml

          1. Coating culture flasks with monomeric Collagen A
          The Collagen molecules remain monomeric in solution at a pH below 3.8 and lower ion concentration, there is no fibril formation.
          We recommend two different coating systems, depending on when you need the culture flasks: either for immediate use after production or after a longer storage period.

          To use the culture flask immediately
          • Mix 1 ml Collagen A (cat. no. L7220) with an equal volume of ultra-pure water (cat. no. L0015, L0020) or with 1x PBS (cat. no. L1815), adjusted to a pH of 3.0 - 3.5. The PBS solution (cat. no. L1815) is adjusted to a pH of 3.0 - 3.5 as follows: Add 1 ml 1 N HCl to 100 ml PBS solution.
          • Add 2 ml of the diluted Collagen solution per 10 cm2 area of culture flask and incubate for 2 - 3 hours or overnight at room temperature (≤ +25°C) (alternatively for 30 min at +35 - +37°C). Remove solution and wash with ultra-pure water or with 1x PBS (pH approx. 7.3). Use the culture flask immediately.
          (1 ml Collagen A/10 cm = 100 µg Collagen/cm)

          To store culture flasks for a longer period
          • Mix 1 ml Collagen A with an equal volume of ultra-pure water (cat. no. L0015, L0020).
          • Add 2 ml of the diluted Collagen solution per 10 cm2 area of culture flask or 20 µl per well of a microtitre plate.
          • Incubate overnight at room temperature (≤ +25°C) in the air stream of a Laminar-Airflow system and let the coating solution dry to form a collagen film. The culture flasks can be stored sterile packed until use (cool, dry storage). Before use, rinse briefly with culture medium.

          2. Coating culture flasks with fibrillar Collagen A
          Collagen fibril formation is carried out at a pH above 4.0, in particular at physiological pH 7.0 - 8.0, at physiological ion concentration and warming to a maximum +37°C.
          We recommend two different coating systems for fibrillar Collagen A, depending on when you need the culture flasks: either for immediate use after production or after a longer storage period.
          To produce the neutralised Collagen solution cool all the solutions in an ice-water bath at approx. +2 - +8°C and mix.

          To use the culture flask immediately
          • Mix 2 ml Collagen A with 0.2 ml PBS solution (10x) (cat. no. L1835).
          • Adjust the pH to pH 7.3 ± 0.2 by adding approx. 10 µl 1 N NaOH.
          • Add 2 ml of solution per 10 cm2 area of culture flask and incubate for at least 3 hours or overnight at room temperature (≤ +25°C) (alternatively for at least 60 min at +35 - +37°C).
          • Remove solution and wash with 1x PBS (pH approx. 7.3) or with nutrient solution. Use the culture flask immediately.

          To store culture flasks for a longer period
          • Mix 1 ml Collagen A with an equal volume of 0.3 M NaCl solution and with 1 N NaOH solution, adjust to a pH of 7.0 - 8.0.
          • Add 2 ml of the diluted Collagen solution per 10 cm area of culture flask or 20 µl per well of a microtitre plate.
          • Incubate overnight at room temperature (≤ +25°C) in the air stream of a Laminar-Airflow system and let the coating solution dry. The culture flasks can be stored sterile packed until use (cool, dry storage). Before use, rinse briefly with culture medium.
          Specifications:

          Sterility: Pass
          Cell Attachment: Pass
          pH: 1.8-2.4
          Osmolality: 0.01-0.05 Osm/kg H20
          Collagen Content: 0.09-0.15%
          Hydroxyproline: 0.012-0.019%
          Color: Colorless
          Appearance: Clear

          Catalogue NumberL7220-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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