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Can mycoplasma grow outside of a cell?

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.

How to determine if a compound interferes with Toxilight Non-Distructive Cytotoxicity BioAssay

#1-2 is to determine if the media contains Adenylate Kinase (AK), the enzyme detected in Toxilight™ Assay1. Run compound in buffer with ToxiLight™ Reagents.There should be no light output.2. Run compound/Media with ToxiLight™ Reagents.If...

Which cell numbers should I use when running the ToxiLight Non-Destructive Cytotoxicity BioAssay

Toxilight works much better at low cell numbers, for adherent cell lines < 10,000 in one well of a 96well plate 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...

What is HEPES? Why is it added to cell culture media?

HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid ) is a zwitterionic organic chemical buffering agent; one of the twelve Good's buffers. HEPES is widely used in cell culture, largely because it is better at maintaining physiological pH...

Can I use the ViaLight Plus Assay also, if my cells are cultured in 24well plates, 6well plates or 12 well plates?

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 µl culture use 50 µl cell lysis). Wait 10 minutes for the lysis . Then follow the protocol...

Do you know if treating cells with 10mM MgCl2 would interfere with the ToxiLight Non-Destructive Cytotoxicity BioAssay?

Magnesium (Mg) is a cofactor for both luciferase and Adenylate Kinase (AK) and will speed up their reactions, but this amount of MgCl2 wouldn't make a significant difference.

What is the purpose of EDTA in trypsin? 

EDTA is a chelator of metal ions like calcium and magensium. It is used with trypsin because it helps bind up the divalent cations in the medium. It helps to keep the cells from sticking to each other.  If its not there….you would need a higher...

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...

I am using the acute leukemia HL-60 cell line for CAR-T cells experiments. Do you have a transfection protocol for genome editing?

The HL-60 cells are often used as a model for in vitro research for treatment of AML (acute myeloid leukemia) in combination with modified primary human CAR- T cells (T cells expressing chimeric antigen receptors).Transfection protocols can be found...

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...
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