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HomeWestern BlottingWestern Blot Analysis of Immunoprecipitation (IP-Western)

Western Blot Analysis of Immunoprecipitation (IP-Western)

IP-Western analysis remains a popular technique for identifying protein-protein interactions and identifying unknown proteins in a multi-protein complex. The steps include cell lysis, formation of the antibody-antigen (immune) complex, precipitation of the immune complexes, and analysis by Western blotting.

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High Background

High background, leading to nonspecific bands on Western blot

Figure 1.High background, leading to nonspecific bands on Western blot.

No Target Protein Eluted, or Not Enough Target Protein Eluted

Extra Band(s) at 50 kDa and 20 kDa on Final Western Blot that May Mask Band of Interest

In a cross-linking IP western, the heavy and light chains of the primary capture antibody are excluded from the sample.

Figure 2.In a cross-linking IP western, the heavy and light chains of the primary capture antibody are excluded from the sample.

Guide to Choosing Immunoprecipitation Beads

Relative Affinity

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