for Mouse Anti-Human MCP-1*:
The following protocol used formalin-fixed paraffin-embedded
human tonsil tissue (supplied by the Alfred Hospital, Australia).
and rehydrate the tissue section.
heat-induced antigen retrieval with 1mM EDTA pH 8.0 at 123˚C for 3 minutes
in a Pascal pressure chamber (Dako).
Wash the slide twice for
three minutes each (50mM Tris-HCl pH 8.0, 150mM NaCl in distilled water).
the tissue section with 1% BSA/PBS block.
the tissue section overnight at 4˚C with Mouse Anti-Human MCP-1 at 10.0 μg/mL. Wash the slide twice for three minutes
- Incubate the tissue section with the
Envision+ HRP polymer kit (Dako) according to the manufacturer’s protocol. Wash the slide twice for three minutes
the tissue section with the DAB chromogen.
the tissue section with hematoxylin.
the sections with DPX mounting medium and image sections with an Olympus
Provis light microscope (Monash Micro Imaging, Monash University,
* Protocol and
staining provided by Dr. Lauren Binge, Laboratory of Prof. Charles Mackay,
Monash Univeristy, Australia.