Figure 2

Addition of blocking reagents greatly improves signal specificity for peroxidase-conjugated anti-hapten antibodies. Planarians were hybridized with DIG-, DNP-, or FAM-labeled anti-sense RNA probes to Smed-histoneH2B (as indicated) and then blocked in buffer with either 5% horse serum alone (A, B, and C), or in 5% horse serum with 0.75% casein (D, E, and F), 0.75% PerkinElmer Blocking reagent (PEBR) (G, H, and I), or 0.75% Roche Western Blocking reagent (RWBR) (J, K, and L) prior to incubation with the appropriate peroxidase-conjugated anti-hapten antibody in the same blocking buffer. Images are maximum intensity projections of planarian heads imaged using identical settings and false colored in ImageJ using the fire look up table. Scale bar: 100 μm.