Hi,

I am planning on using a MTT-based assay for assessing cell proliferation/viability. Has anyone used the alternatives XTT or WST-1? If so, which of the three dyes is best?

Yes, I have used all three. WST-1 is the most sensitive of all three. It is supplied in a liquid form. If your choices are between the MTT and XTT--go with the XTT. You can save some steps with the XTT because it has been formatted so that the color change occurs while proliferation occurs. You will not need to add DMSO for the color change as you would need to do for the MTT.

Hai
I have done MTT, if u want i could help u vth MTT. just PM me

[quote=vasipalli]Hai
I have done MTT, if u want i could help u vth MTT.just PM me. Thanks but I have already completed the MTT assays. Instead of using the WST1 I used the MTT assay.

Many thanks for the offer of help.

I want to do MTT assay for M NFS 60 cell lines for GCSF. I tried doing with 70,000 cells /ml after 48 hours treated with MTT and after 24 hours treated with acidified SDS but i didnt get gradation in OD ie all the OD are equal for 3 subsequent 10 fold dilution. I dont know why
Pls help me in solving this problem
Thanks and regards
Nazia Sulthana

vasipalli said:

Hai I have done MTT, if u want i could help u vth MTT. just PM me

want to do MTT assay for M NFS 60 cell lines for GCSF. I tried doing with 70,000 cells /ml after 48 hours treated with MTT and after 24 hours treated with acidified SDS but i didnt get gradation in OD ie all the OD are equal for 3 subsequent 10 fold dilution. I dont know why
Pls help me in solving this problem
Thanks and regards
Nazia Sulthana
[quote=RockyDoc][quote=vasipalli]Hai
I have done MTT, if u want i could help u vth MTT.just PM me. Thanks but I have already completed the MTT assays. Instead of using the WST1 I used the MTT assay.

Many thanks for the offer of help.[/quote]

Reduce cell numbers - ideally you should titre it. 70 k of those in a 96 well plate is very crowded.
Also, are you actually doing the MTT assay for 24 h?? That will virtually guarantee saturation - 3-6 h is a standard time (treatment with GCSF can be as long as you want, but to get the most info out of your expt, you should have a kinetic assay - 12, 24, 36,48 h of GCSF, followed by 3-6 h of MTT, followed by acid alcohol/acid SDS) - you would ideally need to do this first before titering the GCSF.

i need help with mtt assay as wanna know about the procedure and the fullform of it..i also need somthing on apoptosis kita

what cells are you working with and which apoptosis kit ? Do you want to quantify apoptosis or is it enough to just detect if there is apoptosis occurring or not ?