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What is the effect of pH on the Luciferase-based BioAssays (ViaLight™ Plus, MycoAlert®, ToxiLight®) Lonza is offering?

A low pH can certainly affect the luciferase system used in all of these products. The luciferase will not work if the pH is below 6.5 and performance will rapidly decay below pH 7.2. The reagents in these kits will buffer back to some degree but...

How can cells be cured of mycoplasma contamination?

Where contamination has occurred, and the sample absolutely cannot be discarded, the MycoZap™ Mycoplasma Elimination Reagent has been optimized to eliminate mycoplasma with minimal toxic effects on the cells.The MycoZap™ Reagent eliminates mycoplasma...

Do you have a protocol for stimulation of mouse B cells?

Yes, if you want to transfect mouse B cells, we recommend to stimulate the cells before. The information is also available in the mouse B cell Nucleofection Protocol. Our R&D has used following combination : RPMI1640 supplemented with...

How much sample do you need for accurate results with MycoAlert® and MycoAlert® PLUS Mycoplasma Detection Kits? Do you need cells or just supernatant?

The standard protocol calls for taking 2 ml of culture media, spinning down any cells and removing 100 µl of supernatant. Smaller initial aliquots of media may be taken to start with if necessary.

What is the principle behind the ViaLight™ Plus Cell Proliferation and Cytotoxicity Assay?  

ViaLight™ Plus BioAssay Kits use bioluminescence catalysed by the firefly luciferase enzyme to measure intracellular ATP.  Since ATP is central to the metabolism of all cells, the intracellular concentration is very precisely...

Do intracellular 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...

Can the ViaLight™ Plus 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...

What instrumentation is needed to run the ViaLight™ Plus 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...

What media should be used for spheroid formation with hepatocytes?

Lonza's QC uses HCM (CC-3198) supplemented with 20% FBS and 25 mM Hepes. No plating media is necessary when seeding for spheroids. 

What factors interfere with the ViaLight™ Plus Assay?

Certain compounds will quench the light emission in bioluminescent reactions.  Typically these are strong red dyes (however, the ViaLight ™ Plus Assay has been designed to function in all phenol red containing media commonly used in...
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