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Do you have any tips for the culture of primary cells or cell line prior to Nucleofection?

Yes. Please see our Techincal Reference Guide which contains many helpful tips.You can find additional information in the Optimized Nucleofection Protocol for your cell type of interest.

What is the effect of cellular ATPases on the results obtained with the ViaLight Plus Cell Proliferation and Cytotoxicity BioAssay?

The ViaLight™ Plus Cell Lysis Reagent inhibits ATPase activity - mainly by using a low pH and denaturing the cellular proteins. This is why the ViaLight™ Plus Assay has such a good half life. So cell expressing high ATPase will probably not...

How can I run a standard curve for ToxiLight Non-Destructive Cytotoxicity BioAssay?

ToxiLight does not need a standard curve. You can just plot the RLUs produced including an untreated control. If you want you can create a 100% lysis control by using the ToxiLight™ 100% Lysis Control Set LT07-517 or by heating a...

Does phenol red in the cell culture medium interfere with the ToxiLight Non-Destructive Cytotoxicity BioAssay?

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...

Do all the luciferase based reagents in the Lonza BioAssays need to be protected from light?

Yes, all of our reagents include in ToxiLight™ Non-Destructive Cytotoxicity BioAssay, the ViaLight™ Plus BioAssay, the MycoAlert™ and MycoAlert™ Plus Assay, the PPiLight™ Inorganic Pyrophosphate Assay and the PDELight™ HTS cAMP Phosphodiesterase...

I tested my cell culture supernatant negative for mycoplasma contamination using the MycoAlert® Mycoplasma Detection Kit or the MycoAlert® PLUS Mycoplasma Detection Kit. But when I tested the same cells using a PCR-based Assay, I obtained a positive result.

It is possible that the PCR is picking up dead mycoplasma whereas the MycoAlert® and MycoAlert® Plus Assay will pick up only viable mycoplasma. It is possible of course that there is contamination in the PCR giving a false positive.

Can cell free supernatants ( NOT lysed) be stored for later testing with ViaLight Plus BioAssay?

For supernatants there will be exogenous ATPases present that will degrade ATP so it should really be treated as if it were cells. Freeze as quickly as possible and store at -80C, for no more than 4 weeks. The ATP will be OK with the freezing.

For Lonzas MycoAlert® Mycoplasma Detection Assays is it possible to convert the B/A value of positive control included in the MycoAlert® Assay Control Set to CFU of mycoplasma?

The MycoAlert® and MycoAlert® PLUS Assay are not quantitative so you cannot relate the ratio to an actual value. Different mycoplasma species will give different ratios due to different levels of expression of the enzymes we measure -...

What are inhibitors of the MycoAlert® Mycoplasma Detection Kit

The main inhibitors of the MycoAlert® Reaction would be things that affect the luciferase detection. Agents which potentially quench the light signal include high levels of salt, EDTA and other chelating agents, very high levels of sera,...

Do you have recommendations for the type of samples used for the MycoAlert® Mycoplasma Detection Kit and the MycoAlert® PLUS Mycoplasma Detection Kit? And do you have recommendations related to sample storage?

The MycoAlert® Mycoplasma Detection Kit and the MycoAlert® PLUS Mycoplasma Assay require the use of cell free supernatant only! Cells MUST be removed at 200xg for 5 mins.For optimal assay performance, supernatant should be tested...
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