Hello everyone,
To work with a neural cell type, I will cover the flasks and dishes I use with poly-l-ornithine. I have a Sigma product with catalog number P3655. In the datasheet, I was unable to find any information about the storing conditions for solution aliquots. Can I aliquot the stock I have prepared 10 mg/ml in water? Is there any problem if I freeze-thaw these aliquots? Or can it be stored at +4 after being prepared? Also, in the specification, a 0.22 PTFE filter was suggested; has anyone ever used it? Will filtering my final product cause any adsorption issues?
Thank you very much.
Hello Gülşah Torkay
You may aliquot the stock (10mg/ml) prepared in sterile water and store the aliquots at -20 degree C. Avoid multiple freeze-thaw cycles to maintain product integrity. So aliquots should be prepared in such a way that they may be used for one time or max. two times. The optimal coating concentration is determined by the end user. Generally, the coating concentration range from 10ug/ml to 100ug/ml which is based on cell type and application.
Regarding filtering the final product, you need to follow the instructions provided in the specification. The final product need to be sterile filtered through a 0.22 µm filter using a teflon filter (PTFE filter) which minimizes any adsorption to the filter.
Best.
Hello Malcolm Nobre ,
Thank you for your answer. Unfortunately, the PTFE filter did not work. I think my filter was hydrophobic.
Hi Gülşah Torkay ,
I regularly store PLO in aliquots for both cortical and motor neuron cultures.
I use a Pall 4614 low binding filter. I usually have no issues with filtering through these. Something you may need to use two or three if you are filtering large volumes of PLO solution.
I then aliquot in to 1mL sterile tubes at 1mg/mL, and store in the -80 for up to 6 months, and have never had issues. Use at your desired concentration once thawed, avoiding any freeze thaw cycles.
Hope this helps!
Chris
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