Hi Nadine, 2 things I like to recommand. 1. incubate the primary ab at room temperature becaus higher temperature forces the antibody binding. 2. run a dilution raw of the primary abs to optimize the result. May be you have to use higher concentrations of the primary abs. I am working with GAD67 from Millipore (MAB5406) in a dilution 1:100 in 0,2 % Tx in TBS and 1% normal goat-serium over night at room temperature . And I detect this ab with Goat-anti-Mouse-CY2 1:200 for 90 min at rt.

If the results are not suficient you should use more sensitiv detection systems like Streptavidin labeled with Fluochromes in combinatoin with biotinylated secondary abs. But even in this case you have to titrat the optimal dilution of the primary abs. Good luck!

Hi Nadine. I hope you will share your experience with these antibodies on www.pAbmAbs.com. Here, scientists can share information about the antibodies they use in their research, and your experience will be crucial to some other scientist looking for suitable antibodies 

Alternatively you can also look for other antibodies for your targets that has been validated by scientists within your filed. I have earlier used another antibody against GAD67:

http://pabmabs.com/wordpress/?p=1498

This works excellent for IHC. If you have the option to use this instead i would highly recommend this. 

As a bonus i can tell you that all reviews received on www.pAbmAbs.com anter the lottery for €500 for personal spending. Every review counts as a lottery ticket. 

I hope this helps you.

Kind regards,

Is these perfused or frozen sections (postfixed with PFA) you use? Gad67 ABs do not work very well on frozen sections.

The Millipore antibody works on PFA fixed free floating sections as well as on non fixed cryosections. The native cryosections should be fixed in 4% PFA over night in the fridge before you start with your immuno.