Best substrate for Western-HRP

WSchick at AOL.COM WSchick at AOL.COM
Thu Jun 17 07:45:22 EST 1999


In a message dated 06/17/1999 3:06:34 AM Pacific Daylight Time, 
a8803349.nospam at unet.univie.ac.at writes:

>  the best for chemilum.  Western-HRP
>  detection. I am currently using ECL (Dupont), but I am not very
>  satisfied, because it has a short expiry period and sometimes gave
>  only very fade bands (everything else was OK.)

Pierce has a long-lived Super Signal Dura substrate.  There are three 
cocktails.  A standard for film,  a Dura for Chemiimaging, and a Femto? for 
ultra sensitivity.  I've used mostly the Dura which used to be called Super 
Signal Ultra if you check any references.

You can get a free 20ml DURA sample for Chemiimaging from 
info at alphainnotech.com who recommend it for their Fluorchem and ChemiNova 
digital camera imagers. I am not sure an Austrian distributor is listed on 
their web site, but the samples are available from any Alpha Innotech 
distributor or from Alpha Innotech directly. 

The sample also works for film. The substrate works for Northern and Southern 
blots if you have an HRP labeled probe or biotinylated HRP in your protocol.

Try the sample with only one change of your procedure:  dilute your second 
antibody to 200,000:1 to avoid burnout of the image.  Once you try it, you 
can then optimize your first antibody also--there can be a significant 
reduction in first antibody use with Dura.

You can also ask alpha innotech for a paper that compares ECL and dura for 
chemi imaging.

Walt Schick



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