ELISA technical guide and protocols

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TECH TIP # 65 ELISA technical guide and protocols Table of Contents Introduction ........................................ ........................................ ........................................ ........................................ .................2 Variations between ELISA protocols ........................................ ........................................ ........................................ ..................3 A. Antigen B. Direct vs. Indirect Detection of C. Biotin Signal Amplification ........................................ ........................................ ........................................ ..................3 D. Fluorescence ........................................ ........................................ ........................................ ........................................ ..4 E. Enzymatic Detection.

When multiplexing using labeled secondary antibodies it is essential to use detection antibodies from different species in order to distinguish the separate signals. The detection limit is typically around 100pg/well which is less sensitive than colorimetric or chemiluminescent detection.

  Multiplexing