Western Blotting – The principles and a comparison of indirect vs. direct immunodetection

Western Blot - Lightning-Link HRP table

Western blotting is a powerful and commonly used tool to identify and quantify a specific protein in a complex mixture. Although Western blotting is simple in principle, for a successful outcome, scientists must be aware of the technical caveats which must be overcome.

Topics discussed in the webinar include:

  • Sample preparation
  • SDS-PAGE
  • Western Blot transfer
  • Blocking
  • Indirect Detection – primary antibody incubation and indirect detection
  • Indirect Vs. Direct antibody detection – advantages and disadvantages
  • Direct detection for multiplex fluorescent Western blotting
  • Problems with sourcing labeled antibodies for direct detection
  • The solution simple antibody labeling kits
  • Questioning the secondary antibody amplification hypothesis

At Innova we aim to help you generate optimum antibody conjugates for use in your application. With this in mind we developed our range of Lightning-Link® conjugation kits: quick and easy to use, with no need of separation steps, they are ideal for western blotting (and many other applications).

Please contact us for more information on antibody conjugation kits and their use in western blotting applications.