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. 2019 Jan 10;67(4):275–289. doi: 10.1369/0022155418824092

Figure 4.

Figure 4.

Effect of analyte concentration on ER immunostaining using the ER 1D5 MAb, with and without HIER. It is presented as a contrast to the data of Fig. 2, showing dependence on HIER at all analyte concentrations. Panels A and B depict immunostaining of peptide epitopes (coated onto glass microbeads) with (A) and without (B) HIER. These microbeads bear an average of 934,651 ER peptides per microbead. In Panel C, the ER peptide concentrations per microbead are listed above the vertical bars. Panels D and E depict immunostaining of the same peptide epitopes with (D) and without (E) HIER, except that these microbeads bear an average of 8187 peptide epitopes per microbead. The 1331 molecules per microbead group is not shown because it is below the limit of detection regardless of HIER. The data (Panel C) represent the mean ± SD of triplicate slides. Scale bar, 10 µm. Abbreviations: ER, estrogen receptor; HIER, heat-induced epitope retrieval.