Posted 19 May 2010 - 01:16 AM
currently i am perforng mtt assay but i am facing problem in getting any consistent result in finding out ic50 value of the compound.i am getting same od for my blank and the wells with compounds.wavelength that i am using is 490 nm.please someone help me.
Posted 19 May 2010 - 06:05 PM
Posted 19 May 2010 - 11:48 PM
Posted 20 May 2010 - 12:22 AM
Thanks for the reply. ya i know that but problem is that we do not have 570 nm wavelength (lack of facility) and i was also thinking may be because i am not using 570 nm that's why i am not getting result.but i did my optimisation at this wavelength and result was ok but when i started doing drug screening then problems started.
even if u got any result i wud anyways never rely on that... the detection wavelength is different.. yiou cannot scientifically prove any argument right using this assay system!!!! look for a system were u can read at 570...!!!
moreover 490 adn 570 dont even fall in the same visible color spectrum range.... so its a big no no from my side!!!
sorry if this is not encouraging... but i wont rely on such data!!!
Edited by Prep!, 20 May 2010 - 12:24 AM.
Posted 20 May 2010 - 12:51 AM
if i go for 565 nm that will be ok or not??
yeah tat is still much more better than your previous wavelength!!!! as the bandwidth of a 570 filter will anyways allow 565 nm light to pass!!!
Best luck!!! and as bob suggested it wud be good if u can have a reference wavelength at 630..
Posted 20 May 2010 - 02:05 AM
Posted 20 May 2010 - 02:13 AM
that's great i'll try this and let you know any change in my result. thanks a lot for suggestions to everyone.cheers !!!!
Best luck oasis!!!
Posted 20 May 2010 - 06:07 PM
I am guessing measuring at 490 didn't work as it probably is not a part of the spectrum that is linearly related to the amount of formazan produced.
Posted 21 May 2010 - 01:20 AM
Posted 25 May 2010 - 09:53 PM