test for the quality of my secondary-HRP conjugated ab - (Aug/24/2005 )

Hi,

I suspect that there is a problem with my secondary antibody...I use National Diagnostic HRPLA substrates A and B...I want to see if there is a problem....does anyone have any idea??

thanks sooo much

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You can mix the ECL reagent A and B with a little secondary Ab (1 micro liter) in a microtube. If your secondary Ab is okay, the mixture will generate the fluorescence in the dark.

Good Luck!

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I would suggest that you spot the secondary on a memebrane and then do the ECL. If you get a spot on the film (or monitor if your using a gel doc system) then it still works.

saltycookie provided an interesting idea. I've never tried it but I'm not sure how much signal you will see with your eyes if you do the ECL in an eppendorf.


Good luck!

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I have on occasion done what saltycooky has done.

Its quick and sleazy and provides the answer.

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