Brandtech Hydrograde plates Blocking Buffer

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Brandtech Hydrograde plates Blocking Buffer

Postby ryanmichaeljohnson » Sep 28 2015 7:51 pm

Does anyone have a recommendation for a blocking buffer for BrandTech's 96 well hydrograde plates?

I have tried a variety of different pHs for 1%-5% BSA and NFDM. Next I will try horse serum and Casein. I am waiting on Fish Gelatin to arrive.

Do you have any other ideas?

I am attaching my monoclonal antibody in 0.1 M Sodium Bicarbonate pH 9.6 overnight at 4C on shaker.

Thanks,

Ryan
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Re: Brandtech Hydrograde plates Blocking Buffer

Postby mdfenko » Sep 29 2015 11:50 am

horse serum may work if your secondary antibody is made in horse. if not then serum from the species in which the secondary is made should be used.

in general, nfdm should not be used with avidin-biotin systems. casein should not be used if trying to detect phosphorylated proteins.

sometimes a mixture of blocking agents are effective. we use bsa and normal serum for most of our elisas.

hydrograde plates are hydrophilic and bind glycoproteins well. you can try a glycoprotein that doesn't react with your antibodies (eg collagens).
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