Tissue samples were fixed with 10 % buffered formalin and 6-μm-thick tissue sections were cut. The sections were then incubated with monoclonal antibody (for PIWIs, 1:100; CDKs, 1:200; Cyclins, 1:200) and the same secondary antibody that was used for western blotting. Horseradish peroxidase streptavidin complex (Beyotime, Wuhan, China) was used to visualize the signals. Sections were color-developed with diaminobenzidine and stained with hematoxylin (Beyotime).
Horseradish peroxidase streptavidin complex
Manufactured by Beyotime
Horseradish peroxidase streptavidin complex is a protein-based conjugate that combines the enzymatic properties of horseradish peroxidase with the high-affinity binding of streptavidin to biotin. This complex is commonly used as a detection and labeling reagent in various biological and biochemical applications.
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2 protocols using horseradish peroxidase streptavidin complex
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Immunohistochemical Analysis of Proteins
2
Immunohistochemical Analysis of Tissue Sections
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The samples were fixed with 10% buffered formalin and embedded in paraffin. Tissue blocks were cut into 6-µm sections. The sections were blocked with normal rabbit serum (Santa Cruz Biotechnology, Inc.)and then incubated with the same monoclonal antibodies as those used for western blotting (1:100 dilution for STAT6 and Bcl-xL) at 4° C overnight. After washing with TBST, the sections were stained with the horseradish peroxidase streptavidin complex (Beyotime Institute of Biotechnology, Inc.). The sections were color-developed with diaminobenzidine (Beyotime Institute of Biotechnology, Inc.). The sections were stained with hematoxylin and observed under a Leica 400 light microscope (Leica Microsystems, Inc., Buffalo Grove, IL, USA).
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