Hello everyone!
I am just about to test my cell culture samples to determine the cell viability via the glucose assay kit.
However, since the assay was designed to measure the glucose content in the cell culture medium to determine the glucose concentration, is there any specific requirement of how much of the volume of DMEM should be used to test and how many cells should be taken into account when I am testing the samples over time? When should I start to test (like whatever percentage of confluence)? I know this may confuse some of you, but what I am interested in is purely is there any volume/cell number tradition that people usually follow when doing the glucose assay?
Thanks in advance!
I think you are asking scientific community to plan an experiment for you. What your supervisor told you before buying you a kit
Sorry for making you feel this. My advisor is kind of reckless and he just wants me to figure it out.
I searched some papers but they never mentioned how much of the culture medium they used.
I never mean to ask the scientific community to plan any thing for me.
I just want some suggestions. If you could, I really appreciate it.
Thanks,
Hi. I didn't know if you buy a kit or you are try to choose a good one but sometimes it is useful to read the kit datasheet more that the articles methods.
Maybe they are not the kit more useful for you but provide examples of culture media volume and number of cells recommended:
https://www.promega.com/-/media/files/resources/protocols/technical-manuals/101/glucose-glo-assay.pdf?la=en
https://www.abcam.com/ps/products/65/ab65333/documents/ab65333%20Glucose%20Assay%20kit%20Protocol%20v11a%20(website).pdf
Hope that can help you
PS Don't be afraid to ask in particular if you advisor is not enough professional to bother of your training!
Cell density (seeding )should be optimized and uniform throughout the experiment and medium volume should be uniform as well throughout the experiment. For example, in the case of 96 w/p use 200 ul, in case of 12 w/p 1ml and in case of 6 w/p use 2 ml of medium. I hope it may help you.
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