Samples of MCF-7 and TAMR cells were fixed in 3.7% formaldehyde before incubation with pH 8 EDTA buffer in a pressure-cook microwave for 2 min at 950 W for optimal antigen retrieval. This was followed by blocking with serum-free blocking reagent (DAKO) and incubation with mouse monoclonal pZIP7 antibody (MABS1262, Merck Millipore) at 1:8,000 dilution for 1 h at room temperature in a humidity chamber. Slides were then briefly washed with phosphate buffered saline/Tween and incubated with secondary antibody [Mouse Envision labelled polymer-horse radish peroxidase (HRP), No. K4001, DAKO] for up to 1 h. 3’-3’-diamobenzidine (DAB) chromogen-substrate solution (DAKO) was used to visualize target protein and 0.05% (aqueous) methyl green was used to counterstain nuclei. Images were obtained using Olympus BH-2 microscope with multiple fields of view being imaged for analysis.
3 3 diamobenzidine dab chromogen substrate solution
Manufactured by Agilent Technologies
3'-3'-diamobenzidine (DAB) chromogen-substrate solution is a laboratory reagent used for immunohistochemistry and immunocytochemistry techniques. It serves as a chromogenic substrate that produces a brown color reaction when oxidized by an enzyme, such as horseradish peroxidase, which is commonly used as a reporter enzyme in these techniques.
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Lab products found in correlation
2 protocols using 3 3 diamobenzidine dab chromogen substrate solution
1
Immunohistochemistry Optimization for ZIP7 Analysis
2
Immunohistochemistry Optimization for ZIP7 Analysis
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Samples of MCF-7 and TAMR cells were fixed in 3.7% formaldehyde before incubation with pH 8 EDTA buffer in a pressure-cook microwave for 2 min at 950 W for optimal antigen retrieval. This was followed by blocking with serum-free blocking reagent (DAKO) and incubation with mouse monoclonal pZIP7 antibody (MABS1262, Merck Millipore) at 1:8,000 dilution for 1 h at room temperature in a humidity chamber. Slides were then briefly washed with phosphate buffered saline/Tween and incubated with secondary antibody [Mouse Envision labelled polymer-horse radish peroxidase (HRP), No. K4001, DAKO] for up to 1 h. 3’-3’-diamobenzidine (DAB) chromogen-substrate solution (DAKO) was used to visualize target protein and 0.05% (aqueous) methyl green was used to counterstain nuclei. Images were obtained using Olympus BH-2 microscope with multiple fields of view being imaged for analysis.
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