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Many negative bands in WB - (Dec/06/2006 )

Hi all,

I get numerous negative bands in my WB. I know that you can get a negative band as a "burn out" phenomenon when using too much secondary antibody, but in my own experience this burn out phenomenon looks entirely different.

This is what my blot looks like (note that there is a horizontal artefact in the scanned image which I am *not* concerned about, it is probably caused by our scanner):



Does anyone have an explanation for these *many* negative bands?

Thanks very much.

Daniel
Frankfurt, Germany

-Daniel_K-

The only thing that has helped me in the past is diluting out my primary antibody. Or sometimes increasing the amount of protein/lane.

-WAstate-

too much protein per lane or as suggested too much antibody

-The Bearer-

OK, I achieved way better results this week diluting the *secondary* antibody even further... We usually use this particular antibody at 1:1000, but I get good results in my current setting with 1:4000.

Thanks for your comments.

-Daniel_K-

QUOTE (Daniel_K @ Dec 9 2006, 06:33 AM)
OK, I achieved way better results this week diluting the *secondary* antibody even further... We usually use this particular antibody at 1:1000, but I get good results in my current setting with 1:4000.

Thanks for your comments.


Could you please post your new blot so that I can compare the two.

Thanks

-anoopbal-