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IF experiment problem


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#1 nmyimliuxianzhong

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Posted 18 August 2010 - 01:58 PM

Hi
i'm using the Alexa Fluor 568 goat anti-mouse antibody as the secondary antibody to image CSNK1A protein localization with protein per1,I have used the Alexa Fluor 568 goat anti-mouse IgG at dilutions from 1:200 to 1:300 but it never works, i have tried it a lot. i'm sure the CSNK1 and per1 protein express well. Does anyone give me a good suggestion?
Thank you so much!
Leo

#2 laurequillo

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Posted 18 August 2010 - 10:39 PM

Hi
i'm using the Alexa Fluor 568 goat anti-mouse antibody as the secondary antibody to image CSNK1A protein localization with protein per1,I have used the Alexa Fluor 568 goat anti-mouse IgG at dilutions from 1:200 to 1:300 but it never works, i have tried it a lot. i'm sure the CSNK1 and per1 protein express well. Does anyone give me a good suggestion?
Thank you so much!
Leo


I have never use the 568, but with the other secondaries normally the concentration I use is lower. From 1/800 to 1/5000, of course it depends of the antibody, but you could try using less antibody. And you should do a titration of your primary as well
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