Sequential indirect immunofluorescence (seqIF) is a method that enables multiplex spatial proteomics, without the need for highly manual and complex upstream barcoding and conjugation of primary antibodies.
Since seqIF relies on repeated cycles of staining, imaging and antibody elution (i.e. step for removal of antibodies with a dedicated buffer and an optimized protocol) over the same tissue section, high quality results are obtained by (i) optimal elution efficiency on each cycle, (ii) tissue morphology preservation throughout the experiment, and (iii) target tissue epitopes that would remain intact and stable over multiple cycles.
In this technical note, we present data and results showing:
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