I want to use polybrene @6ug/ml for cells. But Sigma Alrich provides it as in 5grams packing. so can any one tell me step wise and in simple language how to dilute it ?
First you need to dissolve it in water or 150mM NaCl to create a stock solution. Make sure you sterile filter this under a hood. In your case, you should make a stock solution at 6mg/mL so that it's an easy 1:1000 dilution when you are adding to your cells. Polybrene is stable for 2 months at 4 degC but it's best to aliquot the rest and freeze at -20 degC.
First you need to dissolve it in water or 150mM NaCl to create a stock solution. Make sure you sterile filter this under a hood. In your case, you should make a stock solution at 6mg/mL so that it's an easy 1:1000 dilution when you are adding to your cells. Polybrene is stable for 2 months at 4 degC but it's best to aliquot the rest and freeze at -20 degC.
Have you experienced degradation of the polybrene stock solution after repeated freeze and thaw cycles? As for example noticing zero infection efficiency of target cells?
Thanks for any comments
Thanks all....I am adding this for those who might find this answers not explicit enough.
To prepare 1mg/ml Polybrene stock, weigh 50mg of the Polybrene powder and dissolve in 50ml 0.9%Nacl solution of Milipore water. This will give you (1mg/ml). Filter sterilize and make aliquots into small tubes. Store at -20 degrees.
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