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What happens if the cells detach in the Quasi Vivo system?

The silicone used int eh system discourages cells from attaching. Therefore, even if the cells are dislodged by the flow, they will not attach elsewhere in the system.

Scale up ViaLight Plus Assay

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 volumein the plate ( ie 100 ul culture use 50 ul cell lysis). Wait 10 minutes for the lysis . Then follow the protocol...

How is the Quasi Vivo system filled? 

We recommend transferring the cell culture inserts from the static well plates to a pre-filled chamber. The remaining media to be used should be transferred into the reservoir bottle. 

Do you know if treating cells with 10mM MgCl2 would interfere with the ToxiLight Assay?

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.

Is the media recirculating or single pass in the Quasi Vivo system?

The system is designed for recirculating media, and we recommend this setup for most applications. Recirculation allows cells to condition the medium with growth factors and signaling molecules, and therefore, improves growth, viability, and the...

Effect of ATPase on ViaLight plus

ViaLight Plus cell lysis reagent inhibits ATPase activity - mainly by using a low pH and denaturing thecellular proteins. This is why VL+ has such a good half life. So cell expressing high ATPase will probablynot affect the ViaLight Plus assay - the...

Standard curve for Toxilight

ToxiLight does not need a standard curve. Normally we just plot the RLUsproduced 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 then...

Is there sufficient cell material to do a Western Blot with the Quasi Vivo system?

This would depend on the protein to be studied. A typical basic system would consist of 2 chambers, each containing between 250K-300K cells depending on the cell types. Highly-expressed proteins such as those in ribosomes should be detectable, but...

ToxiLight Phenol Red

The assay has been developed in the presence of phenol red so all the example data has beenconducted in complete media. Phenol red does quench the light signal so if you compared assays inphenol red free media they would have higher RLUs. The...

Can enzymatic studies be done using the Quasi Vivo system? 

Yes, although the quantity of cell material is needed will depend on the nature of the study, the enzyme, and its relative expression.