I have the roche cell proliferation kit I (MTT). I'm looking at cell proliferation over 9 days. Can I save the samples to do one ELISA at the end or will I have to do them every day?
I moved the ones I had done so far to the fridge yesterday- I came in this morning to see a colour change back to pink- and the readings on the ELISA were much higher?
I would suggest you to change the way you are performing your experiment. What I would do is to seed every day the basal cellular density you are using, and then at day 8 you will be seeding for the last time. On day 9 you will have all the different times from 24 h to 9 days of proliferation. Then you can run everything in one go. I don't recommend you to store the samples. Other possibility is changing to XTT method, which allow you to read multiple times the same sample, but I am not sure if your system will get saturated before you reach the 9th day.
I would suggest you to change the way you are performing your experiment. What I would do is to seed every day the basal cellular density you are using, and then at day 8 you will be seeding for the last time. On day 9 you will have all the different times from 24 h to 9 days of proliferation. Then you can run everything in one go. I don't recommend you to store the samples. Other possibility is changing to XTT method, which allow you to read multiple times the same sample, but I am not sure if your system will get saturated before you reach the 9th day.
I am totally standing with Ernesto. Not only for MTT experiment, I tried some other time -lapse experiments, such as proliferation, cell cycle, confocal, WB; comparing to seed once and harvest everyday, everyday seed and harvest once is much more reliable. It makes sense considering our fixative, or lysate buffer will affect the stability of our protein or epitopes, but we can assume the cells are almost the same during different culture days.
I am afraid not. After you performed MTT assay, the cells were dead already. The phenomena you observed may not be what you want. I suggest you to seed another set of cells to do ELISA.
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