Hi,
I purchased collagen IV powder from Sigma and the website says to dissolve in acetic acid, PBS or water. As I didn't have acetic acid I tried to dissolve it to 1mg/mL in PBS but the collagen has aggregated. Should I let it dissolve at room temperature or in the fridge? Is it still efficient?
Thank you for your help
Collagen IV, like many proteins, can be tricky to dissolve and may require some special handling to go into solution. Although it can be dissolved in various solvents, acetic acid is often preferred because the mild acid helps to protonate the collagen, disrupting its structure and aiding in solubility. That being said, you can also use PBS, but it might take a bit more work.
When dissolving the collagen, avoiding vigorous stirring or high temperatures is important, as these can denature the protein and make it less effective for cell culture. Try these steps:
Remember to sterilize your solution before use. You can do this by passing the solution through a 0.2µm filter.
If your collagen still won't go into solution, it's possible that the collagen has become denatured or is otherwise damaged. You might need to purchase a new batch if you suspect this is the case.
Finally, remember that the effectiveness of collagen coatings can vary depending on the specific needs of your cells. While a coating of 1mg/mL collagen might work well for some cell types, others may require a higher or lower concentration, or a different type of coating entirely. You might need to experiment to find what works best for your specific application.
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