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Do ATP levels differ between different cell types?

Healthy cells in culture contain maximal amounts of ATP and a wide range of metabolic stimuli do not increase these ATP levels.  Any increase in the ATP concentration of a culture can therefore be associated with an increase in the number of...

Could you extract cDNA from the lysed cells after detection using ViaLight?

The lysis of the cells with NRR should not affect the DNA so whilst we haven't conducted this experiment ourselves I cannot see any real reason why it should not be possible to extract the DNA after lysis

Can the ViaLight kits be used for both proliferation and cytotoxicity assay

Yes.  Since the ATP levels are precisely maintained in healthy cells, the level of ATP accurately reflects the number of viable cells in a culture.  In response to a proliferative signal (e.g. a growth factor) the number of cells in a...

Is there any problem using the Vialight Kit due to the presence of immobilized antibodies on the bottom of the microplate?

There should be no problem with the use of the ViaLight kit with immobilized antibodies. You can if you wish confirm this with your own experiment by conducting a control experiment as follows:Coat triplicate wells of a 96 well plate with each...

What instrumentation is needed to run the ViaLight assay?  

To achieve optimal performance and convenience a microplate luminometer should be used.  If reagent injectors are available on the instrument, these should be used since they enable full automation of the assay.  If reagent injectors are...

It appears that the intrinsic luminescence of different cell types varies greatly.  Do you know whether an activated T cell and a resting T-cell harbour the same quantity of ATP?

Yes, activated T cells do have slightly higher ATP levels than resting, however, when you titrate out the cell number there are no significant differences. i.e. the increase in ATP per cell upon activation is nowhere near as great as the effect of...

What amount of ATP is actually in each well?

The sensitivity range of the reaction reported is in moles rather than molar and thus as you are assaying 100ml of your molar solution you will have ten times less ATP when measured in actual moles i.e. your lowest detectable level is actually...

How does the performance of the ViaLight kits compare with other assays routinely used in cell proliferation/cytotoxicity assays?  

We have compared the ViaLight kits to all other assays routinely used in cell viability studies and have found the performance to be superior in every case.  This is due to the combination of extreme sensitivity and wide dynamic range of...

Do we have to use ATP free tips or can we use sterile tips?

When conducting the ViaLight MDA assay I have used Luria Bertani broth for my cultures. It is best if the culture media is ATP free but if it is included as the negative control on the experimental plate then this will give you an idea of the...

Do we have to test the product alone to see if it enhances or inhibits the bioluminescence signal? How can we do to determine the suitability of our products for testing with the ViaLight system?

If you are worried that your product could contribute to the assay if you can add this to 100ul broth and conduct the assay as normal i.e. add 100ul of Bactolyse, wait 5 min and then add 20ul of AMR taking and immediate 1s integrated reading then...
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