Mice were harvested at week 6 for histology. Tissue samples were fixed for 2 h in 4% paraformaldehyde, demineralized for 2 weeks in 16% ethylenediaminetetraacetic acid, and embedded in optimal cutting temperature compound before cryo-sectioning. Tissue morphology was examined with Masson trichrome staining (Thermo Scientific, Waltham, MA). For immunostaining, sectioned slices were treated with blocking buffer consisting of 2% goat serum and 3% bovine serum albumin in 1X PBS and incubated overnight at 4°C in rabbit polyclonal antibody against CD31 (1:100 dilution), osteocalcin (1:100 dilution), or green fluorescent protein (GFP) (1:100 dilution) (all from Abcam, Cambridge, MA). For secondary staining, sectioned slices were incubated for 1 h at room temperature with secondary antibodies: goat anti-rabbit horseradish peroxidase/3, 3′-diaminobenzidine for CD31/GFP (Abcam), and goat anti-rabbit Alexa Fluor 488 for osteocalcin (1:200 dilution) (Invitrogen, Carlsbad, CA). Nuclei were counterstained with Hoechst 33342 stain (Thermo Scientific, Waltham, MA) and images were taken with a Zeiss fluorescence microscope. Sections were stained with all reagents without primary antibody for negative controls.
Rabbit polyclonal antibody against cd31
Manufactured by Abcam
Sourced in United Kingdom
Rabbit polyclonal antibody against CD31. This antibody is designed to detect the CD31 protein, also known as platelet endothelial cell adhesion molecule (PECAM-1), which is expressed on the surface of endothelial cells and some immune cells.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using rabbit polyclonal antibody against cd31
1
Histological Analysis of Tissue Morphology
2
Immunohistochemical Analysis of Tumor Microvasculature
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Histological preparations were prepared from the tumor and the surrounding skin.
Briefly, tissues were fixed with 10% buffered formol (pH 7.0) and paraffin embedded.
Ten-micron-thick sections were cut and stained with H&E. All sections for immunohistochemistry were deparaffinized and rehydrated using graded concentrations of ethanol prepared with deionized water. The slides were transferred into a 0.05 M Tris-based solution in 0.15 M NaCl with 0.1% v/v Triton-X-100, pH 7.6 (TBST).
Endogenous peroxidase was blocked with 3% hydrogen peroxide for 10 min. Antigen retrieval was done using microwave in citrate buffer at pH 6.0. All slides were incubated at room temperature for 1 h with one of two antibodies: rabbit polyclonal antibody against CD31, a validated endothelial marker (Abcam, Cambridge, UK) or monoclonal mouse anti-rat Ki-67, a marker of cell proliferation (DAKO, Ely, UK).
Using a kidney and liver sections as controls, the highest titer of primary antibodies to produce optimal demonstration of microvessels with the lowest acceptable background
Briefly, tissues were fixed with 10% buffered formol (pH 7.0) and paraffin embedded.
Ten-micron-thick sections were cut and stained with H&E. All sections for immunohistochemistry were deparaffinized and rehydrated using graded concentrations of ethanol prepared with deionized water. The slides were transferred into a 0.05 M Tris-based solution in 0.15 M NaCl with 0.1% v/v Triton-X-100, pH 7.6 (TBST).
Endogenous peroxidase was blocked with 3% hydrogen peroxide for 10 min. Antigen retrieval was done using microwave in citrate buffer at pH 6.0. All slides were incubated at room temperature for 1 h with one of two antibodies: rabbit polyclonal antibody against CD31, a validated endothelial marker (Abcam, Cambridge, UK) or monoclonal mouse anti-rat Ki-67, a marker of cell proliferation (DAKO, Ely, UK).
Using a kidney and liver sections as controls, the highest titer of primary antibodies to produce optimal demonstration of microvessels with the lowest acceptable background
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