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Figure 2 | Proteome Science

Figure 2

From: Antibody cross-linking and target elution protocols used for immunoprecipitation significantly modulate signal-to noise ratio in downstream 2D-PAGE analysis

Figure 2

Elution conditions differentially affect elution from different antibodies. Comparison of commonly used protocols to elute target proteins from Dynabeads® Protein A and other bead-based immunomatrices. Elution in hot LDS or SDS buffers generally employed in 1DE protocols results in complete removal of bound proteins (E; eluate, R; residual on beads as monitored by a second round of hot LDS/SDS treatment). In contrast, elution in four commonly used buffers used for downstream 2DE-analysis consistently results in considerable residual target protein remaining on the beads (only residual protein on beads are shown, as monitored by a second elution in hot LDS buffer). Note that the amount of residual protein on the beads is affected both by the elution conditions as well as the type of antibody.

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Proteome Science

ISSN: 1477-5956