Tumor tissues obtained from xenograft models were embedded in paraffin (Sigma-Aldrich) and subjected to IHC. Tissues were deparaffinized and hydrated, followed by permeabilized for 10 min using 0.5% Triton X-100 (Sigma-Aldrich) in PBS. IHC for anti-Ki67 (Abcam) was carried out by the indirect avidin biotin-enhanced horseradish peroxidase method (Vector Laboratories, Burlingame, CA, USA). Sections were observed via a microscope (Nikon, Tokyo, Japan) and assayed using the Image-Pro Premier software offline program (Media Cybernetics, Rockville, MD, USA).
Avidin biotin enhanced horseradish peroxidase method
Manufactured by Vector Laboratories
Sourced in United States
The Avidin biotin-enhanced horseradish peroxidase method is a laboratory technique used for the detection and visualization of target molecules in various biological samples. It relies on the high-affinity interaction between the proteins avidin and biotin to amplify the signal detection of the horseradish peroxidase enzyme.
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2 protocols using avidin biotin enhanced horseradish peroxidase method
1
Quantifying Cell Proliferation in Xenograft Tumors
2
Immunohistochemical Analysis of PHLDA1 in Glioma
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The human glioma tissue array was purchased from Shanghai Outdo Biotech Co. Ltd (Cat# HBraG177Su01; Shanghai, China). The array was depara nized in xylene solution and rehydrated using gradient ethanol concentrations. Antigen retrieval was performed using sodium citrate and the slides were then incubated with H 2 O 2 to block endogenous peroxidases. Thereafter, the primary antibodies PHLDA1 (1:100) was incubated at 4 °C overnight and the signals were visualized by the indirect avidin biotin-enhanced horseradish peroxidase method according to the manufacturer's instructions (Vector Laboratories, Burlingame, CA). After development, all sections were observed by microscope (400×) and quantitative analysis was performed using the Image-Pro Premier software (v.9.0) program (Media Cybernetics, Rockville, MD).
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