The microvessel density (MVD) was detected through immunohistochemistry (the immunohistochemistry kit was purchased from Hoffmann-La Roche Ltd) using CD34 monoclonal antibody. The slices of HCC tissues and adjacent nontumor tissues were taken for routine dewaxing, antigen retrieval, and sealing. After adding CD34 monoclonal antibody (dilution at 1:100; Abcam Co.) of 100 μL into each slice, the tissues were incubated at 4°C overnight. Then, the slices were washed with PBST and added with the biotin-labelled secondary antibody (dilution at 1:500; Abcam Co.) for a 2-hour incubation at 37°C. The streptavidin avidin biotin complex was added, and the slices were incubated at 37°C for 1 h. After DAB color reaction, hematoxylin was applied for a 2-min counterstaining. And the slices were sealed with neutral resin and observed under optical microscope (Olympus) in a low power (×100) for selection of areas with most new vessels. Cell counting was performed in a high power (×400). Each group was assigned with 6 fields, the average of which was calculated as the MVD. The endothelial cells or cell clusters that presented brown color in immunohistochemistry staining and displayed a distinct border with adjacent microvessels and tumor cells in tumor tissues were all new tumor vessels.
Cd34 monoclonal antibody
Manufactured by Abcam
Sourced in United States
CD34 monoclonal antibody is a laboratory reagent used for the identification and characterization of CD34-positive cells. CD34 is a transmembrane glycoprotein expressed on the surface of hematopoietic stem and progenitor cells, endothelial cells, and some other cell types. This antibody can be used in various applications, such as flow cytometry and immunohistochemistry, to detect and analyze CD34-positive cells.
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Lab products found in correlation
2 protocols using cd34 monoclonal antibody
1
Immunohistochemical Quantification of Microvessel Density in Hepatocellular Carcinoma
2
Immunohistochemical Analysis of Tc17 Cells and Microvessel Density
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Paraffin-embedded tissue sections were deparaffinized and rehydrated, followed by microwave antigen retrieval in EDTA (pH 9.1). For immunohistochemical double staining of Tc17 cells, the sections were processed with the Doublestain system (kit-9999), and the following antibodies were used:rabbit anti-human IL-17 antibody (1∶700 v/v; Abcam, USA) and mouse anti-human CD8 antibody (1∶30 v/v; Abcam, USA). For immunohistochemical analysis of microvessel density, the sections were blocked with blocking solution (serum albumin) diluted in phosphate buffer saline (PBS) 3 times for 5 min. Then, the sections were incubated with primary antibody in a moist chamber at 4°C overnight. The CD34 monoclonal antibody (Abcam, USA) was used in this study. After washing 3 times with PBS for 5 min, a horseradish peroxidase (HRP) polymer-linked secondary antibody was added and incubated for 15 min at 37°C. The sections were then visualized with diaminobenzadine (DAB) and counterstained with hematoxylin. Negative control staining was performed with an isotype control and PBS instead of the primary antibody.
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