Epididymal adipose tissues (VATs) of 16–20 weeks HFD-fed mice were mechanically chopped and then digested with collagenase II (Sigma-Aldrich, St. Louis, MO, USA; Cat. No. C2674) for 15 min at 37 °C. After passing cells through a 200 µm cell strainer (VWR, Cat. No. 100490-158) and centrifugation at 1000× g for 10 min, the pellet containing the stromal vascular cell (SVC) fraction was then incubated with red blood cell lysis buffer. SVC single cell suspensions were incubated with fluorescence-tagged antibodies against CD45 (Biolegend, Cat. No. 103116), CD11b (Biolegend, Cat. No. 101206), and F4/80 (Biolegend, Cat. No. 123116). CD45+CD11b+F4/80+ macrophages were purified using SONY MA900 flow cytometer (SONY). In addition, cells were stained with CD11c (Biolegend, Cat. No. 117343) and CD206 (Biolegend, Cat. No. 141715) antibodies to measure the levels of M1 and M2 activation. ATMs were then cultured in IMDM containing 10% exosome-free FBS to produce extracellular vesicles (EVs).
F4 80
Manufactured by Sony
The F4/80 is a lab equipment product designed for the identification and analysis of macrophages in biological samples. It serves as a specific cell surface marker for this immune cell type.
Automatically generated - may contain errors
2 protocols using f4 80
1
Isolation and Characterization of Adipose Tissue Macrophages
2
Isolation and Characterization of ATMs
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(VATs) were mechanically chopped and then digested with collagenase II (Sigma-Aldrich, Cat. No. C2674) for 15 min at 37 o C. After passing cells through a 200 µm cell strainer (VWR, Cat. No. 100490-158) and centrifugation at 1,000
x g for 10 min, the pellet containing the stromal vascular cell (SVC) fraction was then incubated with red blood cell lysis buffer. SVC single cell suspensions were incubated with fluorescence-tagged antibodies against CD45 (Biolegend, Cat.
No. 103116), CD11b (Biolegend, Cat. No. 101206), and F4/80 (Biolegend, Cat.
No. 123116). CD45+CD11b+F4/80+ macrophages were purified using SONY MA900 flow cytometer (SONY). In addition, cells were stained with CD11c (Biolegend, Cat. No. 117343) and CD206 (Biolegend, Cat. No. 141715) antibodies to measure the levels of M1 and M2 activation. ATMs were then cultured in IMDM containing 10% exosome-free FBS to produce extracellular vesicles (EVs).
x g for 10 min, the pellet containing the stromal vascular cell (SVC) fraction was then incubated with red blood cell lysis buffer. SVC single cell suspensions were incubated with fluorescence-tagged antibodies against CD45 (Biolegend, Cat.
No. 103116), CD11b (Biolegend, Cat. No. 101206), and F4/80 (Biolegend, Cat.
No. 123116). CD45+CD11b+F4/80+ macrophages were purified using SONY MA900 flow cytometer (SONY). In addition, cells were stained with CD11c (Biolegend, Cat. No. 117343) and CD206 (Biolegend, Cat. No. 141715) antibodies to measure the levels of M1 and M2 activation. ATMs were then cultured in IMDM containing 10% exosome-free FBS to produce extracellular vesicles (EVs).
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