The dilution of a primary antibody will differ depending on several parameters, such as: 

  • Antibody titer (specific antibody amount)
  • Antigen abundance - Different in Western blot, where the protein is concentrated in a band, compared to a tissue section, where the antigen is spread over a surface. 
  • Detection method - Especially for proteins of low abundance, more sensitive detection methods like chemiluminescnce, combined with fractionation, are recommended. 

The product information sheet, provided with each antibody, contains suggestions regarding initial dilution in which a given antibody can be used. Agrisera antibodies are optimized for Western blot, and the recommended starting dilution is from 1: 1000, up to 1: 100 000, depending on the primary antibody. 

Antibody dilution

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Note: Each technique requires separate optimization, and due to target protein abundance and distribution, more concentrated primary antibodies need to be used in immunolocalization experiments.