Cells were suspended at 1 × 106/mL in 200 µL staining buffer (DPBS + 2%FBS + 2mM EDTA) followed by adding 0.5 µL anti-CD16/32 FcR blocking antibodies (BD, #553142) [45 (link)]. After 10 min incubation at 4 °C additional fluorescently labelled antibodies against the selected markers were added and the cells were further incubated at 4 °C for 15 min. Subsequently, they were washed by adding 1 mL staining buffer per dark 1.5 mL Eppendorf tube and centrifuged for 5 min at 400× g. The resulting pellet was resuspended in 200 µL staining buffer. The DNA intercalating dye 7-AAD (Biolegend, Koblenz, Germany, #420403) was used to exclude dead cells. A weekly calibrated and validated C6 Accuri (BD) or Cytoflex (Beckman Coulter, Brea, CA, USA) flow cytometer was used for acquisition. BD Accuri C6 software (version 1.0.264.21, San Jose, CA, USA) was used for analysis. The following selected antibodies were used (from Biolegend, San Diego, CA, USA): anti-Ly6C-FITC (#128005), anti-CD11b (#101211). Anti-F4/80-R-PE was ordered from Biorad (#MCA497PET). Doublets, debris and dead cells were excluded in all analyses.
Anti f4 80 r pe
Manufactured by Bio-Rad
Anti-F4/80-R-PE is a fluorescently-labeled antibody used for the detection and analysis of mouse F4/80-positive cells, which are primarily macrophages, by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Accuri c6, by BD (2 mentions)
Cytoflex, by Beckman Coulter (2 mentions)
7 aad, by BioLegend (2 mentions)
Anti cd16 32 fcr blocking antibodies, by BD (2 mentions)
Anti ly6c fitc, by BioLegend (2 mentions)
Anti cd11b, by BioLegend (1 mentions)
Anti cd11b, by Bio-Rad (1 mentions)
Rat igg2aκ, by BioLegend (1 mentions)
2 protocols using anti f4 80 r pe
1
Multicolor Flow Cytometry Immunophenotyping
2
Multicolor Flow Cytometry of Myeloid Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were suspended at 1×106/ml in 200 μl staining buffer (DPBS + 2%FBS + 2mM EDTA) followed by adding 0.5 μl anti-CD16/32 FcR blocking antibodies (BD, #553142) [25] (link). After 10 min incubation at 4°C additional fluorescently labelled antibodies against the selected markers were added and the cells were further incubated at 4°C for 15 min. Subsequently, they were washed by adding 1 ml staining buffer per dark 1.5 ml EP tube and centrifuged for 5 min at 400×g. The resulting pellet was resuspended in 200 μl staining buffer. The DNA intercalating dye 7-AAD (Biolegend, #420403) was used to exclude dead cells. A weekly calibrated and validated C6 Accuri (BD) or Cytoflex (Beckman Coulter) flow cytometer was used for acquisition. BD Accuri C6 software (version 1.0.264.21) and Cytexpert 2.0 were used for analysis, respectively. Following selected antibodies were used (all from Biolegend): anti-Ly6C-FITC (#128005), anti-Ly6G-PE (#127607), anti-CD11b (#101211), anti-MHC II-APC (#107613) and anti-CD115 PE (#135505), while anti-F4/80-R-PE was ordered from Biorad (#MCA497PET). Used isotypes (all from Biolegend) were: Rat IgG2c,κ (#400705), Rat IgG2a,κ (#400507) and Rat IgG2b,κ (#400612).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!