The package says 1 mg but the data sheet specifies "1-2 mg/mL in Tris-buffered saline". Any recommendations? I need to prepare coated flasks with 20 ug/mL. Thanks in advance!
Laminin, product number L2020, from Sigma-Aldrich, usually comes in a freeze-dried (lyophilized) solid form. According to the product details, it is recommended to reconstitute the product in 1 mL of sterile phosphate-buffered saline (PBS) or Tris-buffered saline (TBS) to a final concentration of approximately 1-2 mg/mL.
To prepare coated flasks with 20 µg/mL, you would dilute the stock solution accordingly. Here's an example procedure:
Please remember that this is just a general procedure. Depending on your specific experiment, you might need to adjust the concentrations, volumes, or incubation times.
Before starting, it's recommended to double-check the product information or contact Sigma-Aldrich's customer service to confirm the product's form and the reconstitution process. They might have more specific advice for working with this particular product.
Sigma Laminin (L2020) is a lyophilized (freeze-dried) solid product. Therefore, it needs to be reconstituted before use.
The data sheet is likely specifying the recommended concentration range for reconstitution in Tris-buffered saline (1-2 mg/mL), but this can vary depending on the exact application.
To prepare a 20 µg/mL solution, you would first reconstitute the 1 mg of Laminin in an appropriate volume of Tris-buffered saline to get a stock solution (e.g., for 1 mg/mL, you would add 1 mL of Tris-buffered saline). You would dilute the stock solution further to achieve the desired final concentration. For a 20 µg/mL solution, you can dilute the 1 mg/mL stock solution 1:50.
For example, to coat a single well in a 6-well plate, which typically has a surface area of 9.6 cm^2, you might add 1 mL of your 20 µg/mL solution, which would give a coating of 20 µg/cm^2. You would then incubate the coated surface at 37°C overnight for several hours to allow the Laminin to adsorb to the plastic. Afterwards, remove the Laminin solution, rinse the well with PBS, and it should be ready for cell seeding.
Remember to always handle Laminin solutions on ice and with pre-chilled pipettes and tubes because Laminin can degrade quickly at room temperature.
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