Fluorescent measurements-AMCA - (Jan/29/2010 )

Hi all,
I have to measure proteasome activity by measuring fluorescence of degraded substrate Suc-LLVY-AMC. It is recommended to use 380-nm excitation and 460-nm emission filters. But the only available emission filters are 355-nm and 400-nm. Could anyone be so kind to advice on which is better to use, since I have failed to find an info on the subject? 7-Amino-4-methylcoumarin (AMCA) itself has 365-nm ex. Thanks in advance!

-katenkak-

use the 400nm filter. it will block light below 400nm and allow your wavelength through.

-mdfenko-

Thanks!

mdfenko on Jan 29 2010, 05:09 PM said:

use the 400nm filter. it will block light below 400nm and allow your wavelength through.

-katenkak-

Hi all,

I'm a new member of the Bioforum.

Some of you can suggest me how should be better to measure the proteasome activity in the brain tissue?

thanks in advance

-francesca-

do you have an assay for proteasome?

are you asking for a (partial) purification procedure?

-mdfenko-