- Производитель:
- Sigma-Aldrich
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 or with 1x PBS, 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.
To store culture flasks for a longer period:
- Mix 1 ml Collagen A with an equal volume of ultra-pure water.
- Add 2 ml of the diluted Collagen solution per 10 cm2 area of culture flask or 20+D1 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).
- 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 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. The culture flasks can be stored sterile packed until use (cool, dry storage). Before use, rinse briefly with culture medium.
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
biological source | bovine (calf) skin |
Quality Level | 200, |
form | liquid |
packaging | pkg of 6 x 5 mL x |
concentration | 1 mg/mL |
application(s) | cell culture | mammalian: suitable |
shipped in | ambient |
storage temp. | 2-8°C |
WGK Germany | nwg |
Flash Point F | Not applicable |
Flash Point C | Not applicable |
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