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imunofluerecence staining with Alexa 568 dye


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#1 nmyimliuxianzhong

nmyimliuxianzhong

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Posted 19 August 2010 - 03:04 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 Markovnikov

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Posted 19 August 2010 - 03:33 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

What do you mean by it doesn't work? Is everything stained or nothing stained? I'd recommend a longer incubation with your primary, at least overnight, maybe 2 days. make sure you're being careful in your wash steps. Also make sure that your primary is a mouse Ab (or at least the that the goat secondary reacts with it)--seems silly, but an easy mistake to make.




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