Immpact dab peroxidase kit

After following the hydration protocol described above, the tissues were subjected to heated antigen retrieval in Tris/EDTA buffer (pH 9.0), followed by blocking in BLOXALL (30 min) and 10% goat serum in PBS (20 min) obtained from Vector Labs (Burlingame, CA) and detection of rabbit anti-ITGAX (1:100, 4°C, overnight) (Abcam, ab52632; Cambridge, MA) using ImmPRESS^TMHRP anti-rabbit IgG and ImmPACT DAB Peroxidase kits from Vector Labs. The process was repeated for the detection of mouse anti-ITGAM (1:500, 4°C, overnight) (R&D Systems, MAB16991; Minneapolis, MN), employing ImmPRESS^TMHRP anti-mouse IgG (clone #238446) and ImmPACT SG Peroxidase kits from Vector Labs. Tissues were also stained with and without primary and secondary antibodies for both antigens to serve as staining controls.

Slide-mounted anterior ethmoid tissues were sequentially deparaffinized in xylenes and rehydrated in ethanol (Thermo Fisher Scientific; Pittsburg, PA) and ddH₂O, as previously described.30 (link) Tissues were then subjected to heated-antigen retrieval in citrate-based antigen unmasking solution (pH 6.0), followed by antigen blocking with BLOXALL (20 min) and 2.5% goat serum (20 min), all of which were obtained from Vector Laboratories (Burlingame, CA, USA). The tissues were then incubated with polyclonal rabbit anti-human CCR7 (1:200, 4°C, overnight) (Bioss Antibodies, bs-1305R; Woburn, MA, USA), followed by incubation with ImmPRESS^TMHRP anti-rabbit IgG (20 min) and detection with ImmPACT DAB Peroxidase kits from Vector Laboratories (Burlingame, CA, USA). This staining protocol was repeated for monoclonal mouse anti-human CCL19 (1:150, 4°C, overnight) (Invitrogen, OTI2A12; Carlsbad, CA, USA) detection, employing heated antigen retrieval with Tris-based antigen unmasking solution, ImmPRESS^TMHRP anti-mouse IgG (clone #238446), and ImmPACT SG Peroxidase kits from Vector Laboratories.