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This is an area that is up for debate. There is evidence that mycoplasma can survive in the absence of cells, and some may proliferate, but as a rule they are much happier if cells are present.
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#1-2 is to determine if the media contains AK. 1. Run compound in buffer with ToxiLight reagents. There should be no light output. 2. Run compound/Media with ToxiLight reagents. If customer get's light output then most likely there is AK present...
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Toxilight works much better at low cell numbers, for adherent cell lines < 10,000 in 96 and < 50,000 for suspension. We would normally seed 10,000 cells/well when using adherent cells to allow enough room in the well for the control...
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To do the VialIght Plus Assay on larger volume tissue culture ( eg 6 well , 24 well) add the cell lysis reagent equal to half the volume in the plate ( ie 100 ul culture use 50 ul cell lysis). Wait 10 minutes for the lysis . Then follow the...
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Magnesium (Mg) is a cofactor for both luciferase and AK and will speed up their reactions, but this amount of MgCl2 wouldn't make a significant difference.
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ViaLight Plus cell lysis reagent inhibits ATPase activity - mainly by using a low pH and denaturing the cellular proteins. This is why VL+ has such a good half life. So cell expressing high ATPase will probably not affect the ViaLight Plus assay -...
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No, currently we can only offer in-house maintenance services, but we are working on a process for on-site services.
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ToxiLight does not need a standard curve. Normally we just plot the RLUs produced including our untreated control. If they want they can create a 100% lysis control by heating a sample of their untreated cells at 56C for 1h. Measuring this and...
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The assay has been developed in the presence of phenol red so all the example data has been conducted in complete media. Phenol red does quench the light signal so if you compared assays in phenol red free media they would have higher RLUs. The...
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Yes. Please click on the link under related literature.
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