The expression of Notch1 and Fbw7 was detected in histologic sections of tumor xenografts. Sections were cut from formalin-fixed, paraffin-embedded tissue blocks; collected on 3-ethoxy-aminoethyl-silane-treated slides; and allowed to dry overnight at 37 °C. Sections were dewaxed in xylene, rehydrated through graded concentrations of ethanol to distilled water, immersed in 10 mmol/L citrate buffer (pH 6.0), and processed in a thermostatic water bath for 40 minutes at 98°C for antigen retrieval. Anti-Notch1, and anti-Fbw7 antibodies were applied on two slides for each case, and incubations were done overnight at room temperature in a humidified atmosphere followed by a 30-minute incubation of secondary antibody. Slides were then incubated with streptavidin peroxidase and visualized using the 3,3'-diaminobenzidine chromogen (Lab Vision Corp., Fremont, CA).
3 3 diaminobenzidine chromogen
Manufactured by Lab Vision
Sourced in United States
3,3'-diaminobenzidine chromogen is a chemical compound used as a substrate in immunohistochemistry and related techniques. It produces a brown stain upon oxidation, allowing the visualization of target proteins or other biomolecules in biological samples.
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2 protocols using 3 3 diaminobenzidine chromogen
1
Immunohistochemical Analysis of Notch1 and Fbw7
2
Hepatocyte Proliferation Quantification
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The proliferative activity of hepatocytes in liver tissue was evaluated by immunochemical detection of the nuclear non-histone protein Ki-67. Ki-67 staining was performed using rabbit monoclonal SP6 antibody with 3,3′-diaminobenzidine chromogen (Lab Vision Corporation, Thermo Fischer Scientific, Fremont, CA, USA). The mean percentage of Ki-67-positive hepatocytes among total hepatocytes in 10 random 200× fields per animal was used for the Ki-67 labeling index (n = 6/group).
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