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Cd68 percp cy5

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CD68 PerCP-Cy5.5 is a fluorescently-labeled antibody that binds to the CD68 protein. CD68 is a glycoprotein that is highly expressed on the surface of macrophages and monocytes. The PerCP-Cy5.5 fluorophore is attached to the antibody, allowing for detection and analysis of CD68-positive cells using flow cytometry.

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Lab products found in correlation

Siglec f bv421, by BD (2 mentions) Cd64 pe cy7, by BioLegend (2 mentions) Cd115 alexa fluor 488, by BioLegend (2 mentions) Cd93 percp cy5, by BioLegend (2 mentions) F4 80 pe cf594, by BD (2 mentions) Sca 1 allophycocyanin, by BioLegend (2 mentions) Cd16 32 allophycocyanin r700, by BD (2 mentions) Cd11c allophycocyanin 780, by Thermo Fisher Scientific (2 mentions) Cd41 bv421, by BD (2 mentions) Ter119 bv510, by BD (2 mentions) Cx3cr1 bv650, by BioLegend (2 mentions) C kit bv711, by BD (2 mentions) Cd3e buv395, by BD (2 mentions) Cd11b buv737, by BD (2 mentions) Vcam1 alexa fluor 647, by BioLegend (2 mentions) Mhc class 2 alexa fluor 700, by Thermo Fisher Scientific (2 mentions) Tim 4 bv510, by BD (2 mentions) Cd45 bv786, by BD (2 mentions) Cd144 pe, by BD (2 mentions) Ly6g bv605, by BD (2 mentions)

5 protocols using cd68 percp cy5

1

Macrophage Activation and Mitochondrial Dynamics

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Monocyte-derived macrophages were stimulated for 6 hours with 100 ng/mL LPS plus 5 mM adenosine triphosphate (ATP), 3 µM STS, or N gonorrhoeae strain FA1090B at an MOI of 10 or 100. Following stimulation, cells were lifted and washed 3 times in 0.5% FBS/PBS. Cells were stained for CD11b and CD68 expression (CD11b-APC; BioLegend, San Diego, CA, USA and CD68-FITC; BioLegend) for 30 minutes at 4°C and then washed. Prior to analysis, cells were stained with the live-dead stain, propidium iodide (PI; BioLegend) and analyzed on a BD LSR II (San Jose, CA, USA). To evaluate perturbation to the mitochondrial membrane potential, MDMs were stained with Rhodamine 123, a mitochondrial membrane potential dye. Cells were incubated with 1 µM of Rhodamine 123 (Molecular Probes, Eugene, OR, USA), for 30 minutes at 37°C. Following staining, cells were lifted and stained for macrophage markers as described above and were immediately analyzed by flow cytometry. For Rhodamine 123 staining, CD68-PerCP/Cy5.5 and CD11b-APC (BioLegend) were used to delineate macrophage populations.
Ritter J.L, & Genco C.A. (2018). Neisseria gonorrhoeae–Induced Inflammatory Pyroptosis in Human Macrophages is Dependent on Intracellular Gonococci and Lipooligosaccharide. Journal of Cell Death, 11, 1179066017750902.
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2

Comprehensive Flow Cytometry Panel

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The following Abs were used for flow cytometry: CD115 Alexa Fluor 488 (AFS98, BioLegend, 135512), CD93 PerCP-Cy5.5 (AA4.1, BioLegend, 136512), CD144 PE (11D4.1, BD Biosciences, 562243), F4/80 PE-CF594 (T45-2342, BD Biosciences, 565613), CD64 PE-Cy7 (X54-5/7.1, BioLegend, 139314), Sca-1 allophycocyanin (D7, BioLegend, 108112), CD16/32 allophycocyanin-R700 (2.4G2, BD Biosciences, 565502), CD11c allophycocyanin-780 (N418, eBio-science, 47011482), CD41 BV421 (MWReg30, BD Biosciences, 747729), Ter119 BV510 (TER-119, BD Biosciences, 563995), Ly6G BV605 (1A8, BD Biosciences, 563005), CX3CR1 BV650 (SA011F11, BioLegend, 149033), c-Kit BV711 (2B8, BD Biosciences, 105835), CD45 BV786 (30-F11, BD Biosciences, 564225), CD3e BUV395 (145-2C11, BD Biosciences, 563565), CD11b BUV737 (M1/70, BD Biosciences, 564443), CD68 PerCP-Cy5.5 (FA-11, BioLegend, 137010), VCAM1 Alexa Fluor 647 (429, BioLegend, 105712), MHC class II Alexa Fluor 700 (M5/114, Thermo Fisher Scientific, 56532182), Siglec-F BV421 (E50-2440, BD Biosciences, 562681), Tim-4 BV510 (21H12, BD Biosciences, 742774), Live/Dead, fixable UV (Thermo Fisher Scientific, L34962).
Philpott J., Kazimierczyk S., Korgaonkar P., Bordt E., Zois J., Vasudevan C., Meng D., Bhatia I., Lu N., Jimena B., Porter C., Cherayil B.J, & Jain N. (2022). RXRα Regulates the Development of Resident Tissue Macrophages. ImmunoHorizons, 6(6), 366-372.
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3

Comprehensive Flow Cytometry Panel

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The following Abs were used for flow cytometry: CD115 Alexa Fluor 488 (AFS98, BioLegend, 135512), CD93 PerCP-Cy5.5 (AA4.1, BioLegend, 136512), CD144 PE (11D4.1, BD Biosciences, 562243), F4/80 PE-CF594 (T45-2342, BD Biosciences, 565613), CD64 PE-Cy7 (X54-5/7.1, BioLegend, 139314), Sca-1 allophycocyanin (D7, BioLegend, 108112), CD16/32 allophycocyanin-R700 (2.4G2, BD Biosciences, 565502), CD11c allophycocyanin-780 (N418, eBio-science, 47011482), CD41 BV421 (MWReg30, BD Biosciences, 747729), Ter119 BV510 (TER-119, BD Biosciences, 563995), Ly6G BV605 (1A8, BD Biosciences, 563005), CX3CR1 BV650 (SA011F11, BioLegend, 149033), c-Kit BV711 (2B8, BD Biosciences, 105835), CD45 BV786 (30-F11, BD Biosciences, 564225), CD3e BUV395 (145-2C11, BD Biosciences, 563565), CD11b BUV737 (M1/70, BD Biosciences, 564443), CD68 PerCP-Cy5.5 (FA-11, BioLegend, 137010), VCAM1 Alexa Fluor 647 (429, BioLegend, 105712), MHC class II Alexa Fluor 700 (M5/114, Thermo Fisher Scientific, 56532182), Siglec-F BV421 (E50-2440, BD Biosciences, 562681), Tim-4 BV510 (21H12, BD Biosciences, 742774), Live/Dead, fixable UV (Thermo Fisher Scientific, L34962).
Philpott J., Kazimierczyk S., Korgaonkar P., Bordt E., Zois J., Vasudevan C., Meng D., Bhatia I., Lu N., Jimena B., Porter C., Cherayil B.J, & Jain N. (2022). RXRα Regulates the Development of Resident Tissue Macrophages. ImmunoHorizons, 6(6), 366-372.
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4

Flow Cytometry of Tumor-Infiltrating Macrophages

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To prepare single-cell suspensions for flow cytometry, fresh tumor tissue was dissected into ~1–3mm3 fragments and digested with 80 U/mL collagenase (Invitrogen) in DMEM containing 10% FBS for 1 h at 37 °C while shaking. After red blood cell (RBC) lysis, single-cell suspensions were filtered and incubated for 20 min on ice with the following antibodies (1:100): CD45-PE (Cat# 12-0451-81, eBioscience, San Diego, CA), F4/80-PE-Cy7 (Cat# 25-4801-82, eBioscience, San Diego, CA), CD11b-FITC (Cat# 553310, BD Biosciences San Jose, CA), CD206-APC (Cat# 141707, Biolegend, San Diego, CA),CD68-PerCP-Cy5.5 (Cat# 137009, Biolegend, San Diego, CA) for macrophages. Cells were washed with PBS before analysis on the BD LSR-II flow cytometer (Beckman Coulter).
Liang P., Henning S.M., Grogan T., Elashoff D., Ye H., Cohen P, & Aronson W.J. (2022). Effects of dietary omega-3 fatty acids on orthotopic prostate cancer progression, tumor associated macrophages, angiogenesis and T-cell activation—dependence on GPR120. Prostate cancer and prostatic diseases, 25(3), 539-546.
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5

Quantifying Tumor Immune Cell Populations

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To quantify immune cells, single-cell suspensions were prepared from tumor tissue and were incubated with the following antibodies in 1:100 dilution: CD45-PE (hematopoietic cells, eBioscience, San Diego, CA), F4/80-PE-Cy7 (macrophages, eBioscience, San Diego, CA), CD11b-FITC (monocyte/macrophages, BD Biosciences San Jose, CA), CD206-APC (M2 type macrophages, Biolegend, San Diego, CA),CD68-PerCP-Cy5.5 for macrophages (M1 type macrophages,Biolegend, San Diego, CA), CD45-PE, F4/80-PE-Cy7, CD11b-FITC, Gr-1-APC (myeloid cells, BD Biosciences San Jose, CA) for myeloid derived suppressor cells (MDSCs) and Neutrophils, CD45-PE, CD4-FITC (CD4 T cells, BD Biosciences San Jose, CA), CD8-APC (CD8 T cells, BD Biosciences San Jose, CA), B220 (B cells, Biolegend, San Diego, CA) for T and B cells. Cells were washed with PBS before analysis on the BD LSR-II flow cytometer (Beckman Coulter), GFP+ cells were gated for analysis of immune cells as previously described [6] .
Liang P., Henning S.M., Grogan T., Elashoff D., Said J., Cohen P, & Aronson W.J. (2023). Effect of omega-3 fatty acid diet on prostate cancer progression and cholesterol efflux in tumor-associated macrophages-dependence on GPR120. Prostate cancer and prostatic diseases.
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