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mtt assay


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#1 oasis

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Posted 19 May 2010 - 01:16 AM

Hi there,
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.

#2 bob1

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Posted 19 May 2010 - 06:05 PM

MTT is not measured at 490 - read the protocol! Measure at 570 with reference at 630.

#3 oasis

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Posted 19 May 2010 - 11:48 PM

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.

#4 Prep!

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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...!!! :D
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!!! :P

Edited by Prep!, 20 May 2010 - 12:24 AM.

Support bacteria - They are the only culture some people have!!!
Cheers!!!

#5 oasis

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Posted 20 May 2010 - 12:47 AM

HEY thanks a lot for your reply, i will try with 570 nm .

#6 oasis

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Posted 20 May 2010 - 12:49 AM

if i go for 565 nm that will be ok or not??

#7 Prep!

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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.. :)
Support bacteria - They are the only culture some people have!!!
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#8 oasis

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Posted 20 May 2010 - 02:05 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 !!!!

#9 Prep!

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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!!!
Support bacteria - They are the only culture some people have!!!
Cheers!!!

#10 bob1

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Posted 20 May 2010 - 06:07 PM

Yeah, Good luck, hopefully things work for you.

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.

#11 oasis

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Posted 21 May 2010 - 01:20 AM

hi ya again thaks for your reply, in some papers they used 490 nm to carry out MTT assay.Do you really beleve in these papers then?

#12 arera

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Posted 25 May 2010 - 09:53 PM

Please guide us to the paper then... author name, year, etc.

#13 oasis

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Posted 06 June 2010 - 11:40 PM

hi this is the paper which uses 490 nm.

Attached Files

  • Attached File  MTT.pdf   58.99KB   472 downloads





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