For the immunohistochemical staining of aortic root sections, endogenous peroxidase activity was quenched with 3% hydrogen peroxide. Thereafter, the sections were incubated with primary antibodies at 4 °C overnight. Macrophages were detected using rabbit anti-mouse F4/80 antibody (Abcam, Cambridge, UK). IHC-Tek streptavidin-horseradish peroxidase solution (IHC World, Woodstock, MD, USA) and diaminobenzidine substrate (Vector Laboratories, Burlingame, CA, USA) were used to develop the brown reaction product. Slides were counterstained with hematoxylin. Pictures were obtained using a Zeiss confocal microscope. Images were analyzed using ImageJ software.
Rabbit anti mouse f4 80 antibody
Manufactured by Abcam
Sourced in United Kingdom
Rabbit anti-mouse F4/80 antibody is a primary antibody that specifically recognizes the F4/80 antigen, a cell surface glycoprotein expressed on mouse macrophages. This antibody can be used in various immunological techniques to identify and study macrophages in mouse tissues or cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Diaminobenzidine substrate, by Vector Laboratories (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Beyotime (1 mentions)
Paraformaldehyde (pfa), by Beyotime (1 mentions)
Anti asc antibody, by Cell Signaling Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
3 protocols using rabbit anti mouse f4 80 antibody
1
Aortic Root Macrophage Immunohistochemistry
2
Visualizing Dye-anti-CD206 Distribution in 4T1 Tumors
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To investigate the microdistribution of Dye-anti-CD206 in 4T1 tumor tissues, one 4T1 tumor-bearing BALB/c mouse was intravenously injected with 1 nmol of Dye-anti-CD206. At 24 h p.i., the mouse was sacrificed; the tumor was harvested, immediately frozen in optimal-cutting-temperature (OCT) medium, and then cut into 5-μm-thick slices. The tumor slices were incubated with rabbit anti-mouse F4/80 antibody (Abcam, Cambridge, MA) for 1 h at room temperature, and then visualized using fluorescein isothiocyanate (FITC)-conjugated secondary antibody under a confocal microscope. The intratumoral microdistribution of Dye-IgG (as a control) was also determined using the same protocol.
3
Immunofluorescence Assay for Macrophage Detection
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PMs were transfected with CLP/siRNA as described above. After 48 hours, PMs were washed twice with cooled PBS and then fixed in paraformaldehyde (Beyotime Biotechnology, China) for 30 min at room temperature. Subsequently, PMs were blocked with 5% bovine serum albumin (BSA) (Beyotime Biotechnology, China) for 2 hours and incubated overnight at 4°C with anti-ASC antibody (Cell Signaling Technology, USA). The next day, cells were incubated with Alexa FluorTM 594-conjugated goat anti-rabbit IgG (H+L) (Invitrogen, ThermoFisher Scientific, USA) for 1 hour, followed by staining with DAPI (Solarbio LIFE SCIENCES, China) for 10 min. Finally, fluorescence intensity and density were detected by confocal fluorescence microscopy.
Macrophage content in frozen sections of colonic tissue was also detected by IF, as described in previous research.52 (link) The primary antibody was rabbit anti-mouse F4/80 antibody (Abcam, England), and the secondary antibody was Alexa FluorTM 594-conjugated goat anti-rabbit IgG (H+L) (Invitrogen, ThermoFisher Scientific, USA).
Macrophage content in frozen sections of colonic tissue was also detected by IF, as described in previous research.52 (link) The primary antibody was rabbit anti-mouse F4/80 antibody (Abcam, England), and the secondary antibody was Alexa FluorTM 594-conjugated goat anti-rabbit IgG (H+L) (Invitrogen, ThermoFisher Scientific, USA).
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