To induce inflammation in RAW 264.7 cell lines, we treat cells with LPS (lipopolysaccharide) which is supposed to be dissolve in water. Did adding of water to the culture have any effect on cell lines? Can we use PBS instead of water?
Your stock solutions of LPS should be very concentrated. I keep mine at 5 mg/mL. The final working concentration I use for LPS is 1 ug/mL, which in itself is on the high end (low end is usually 1 ng/mL). At that concentration, you would end up effectively adding just 0.2 uL of water (or other solvent) per 1 mL of medium. That amount of water is so insignificant there will be no observable effect by just about any form of measurement.
If you end up making sub-dilutions to make transferring volumes easier, then you should do that in your final working medium (e.g., DMEM).
Dear Raghuram, Basically if you want follow LPS effect, you have to eliminate probable its solvent effect. So, if you dissolve LPS in water as solvent, it is better add equal amount water to wells of the control cells. if , for any reason you need add PBS to the control cells, so dissolve LPS in PBS too.
Finally it may have no effect, displacing water with PBS, but logically for escape from solvent effect, the same solvent is the best choice.
regards
mehdi
Your stock solutions of LPS should be very concentrated. I keep mine at 5 mg/mL. The final working concentration I use for LPS is 1 ug/mL, which in itself is on the high end (low end is usually 1 ng/mL). At that concentration, you would end up effectively adding just 0.2 uL of water (or other solvent) per 1 mL of medium. That amount of water is so insignificant there will be no observable effect by just about any form of measurement.
If you end up making sub-dilutions to make transferring volumes easier, then you should do that in your final working medium (e.g., DMEM).
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