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Ter119 apc

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TER119-APC is a fluorescent-labeled antibody used for the detection and identification of erythroid cells in flow cytometry applications. It targets the TER119 antigen expressed on the surface of erythroid cells at various stages of development.

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Lab products found in correlation

Cd45 apc, by Thermo Fisher Scientific (3 mentions) Fc block, by BD (2 mentions) Cd45 apc cy7, by BD (2 mentions) Cd3 apc, by BD (2 mentions) Cd19 pe, by BD (2 mentions) Cd19 apc, by BD (2 mentions) Cd11b percp cy5, by BD (2 mentions) Cd3 eflour450, by Thermo Fisher Scientific (2 mentions) Cd4 v500, by BD (2 mentions) Nk1.1 apc, by BD (2 mentions) Nk1.1 pe cy7, by Thermo Fisher Scientific (2 mentions) Gr1 apc, by BD (2 mentions) Cd29 fitc, by Thermo Fisher Scientific (2 mentions) Cd31 apc, by Thermo Fisher Scientific (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Facscanto 2, by BD (1 mentions) F4 80 apc, by Thermo Fisher Scientific (1 mentions) Cd11c pe, by Thermo Fisher Scientific (1 mentions) F4 80 pe cy7, by BD (1 mentions) Cd8a percp cy5, by BD (1 mentions)

Spelling variants of this product

Ter119 (107 citations) Anti-mouse Ter119-APC (4 citations) Anti-Ter119-APC-eFluor 780 (2 citations) Anti-TER119-APC (2 citations) APC)–conjugated anti-Ter119 (2 citations)

10 protocols using ter119 apc

1

Flow Cytometry Analysis of Immune Cell Subsets

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SVF pellets were washed with PBS/2% FBS, incubated with Fc Block (BD Bioscience, San Jose, CA, USA) for 15 min and then stained with the following conjugated antibodies (30 min at 4 °C in the dark): CD45-APC-Cy7, CD3-APC, CD19-APC, NK1.1-APC, F4/80-PE-Cy7, CD11b-PerCP-Cy5.5 (all from BD Bioscience), and TER119-APC and CD11c-PE (eBioscience, San Diego, CA, USA) for ATM, CD45-APC-Cy7, CD19-PE, CD4-V500, CD8a-PerCP-Cy5.5, Gr1-APC (BD Bioscience), and CD3-eFlour450, NK1.1-PE-Cy7, TER119-APC, F4/80-APC (eBioscience) for lymphocyte population. After incubation, cells were washed and resuspended in PBS/2% FBS and then analyzed by FACS Canto II (BD Biosciences) and FlowJo software (version 10, Tree Star Inc., Ashland, OR, USA). As described previously [25 (link)], ATM were defined as CD45+, CD3−, CD19−, NK1.1−, TER119−, CD11b+ and F4/80+. T cells (CD19−CD3+NK1.1−), B cells (CD19+CD3−), and NK cells (CD19−CD3−NK1.1+) were gated after excluding the myeloid lineage (Gr1+, F4/80+) and red blood cells (TER119+) among immune cells (CD45+).
Lee Y., Kim Y., Lee M., Wu D, & Pae M. (2021). Time-Restricted Feeding Restores Obesity-Induced Alteration in Adipose Tissue Immune Cell Phenotype. Nutrients, 13(11), 3780.
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2

Multicolor Flow Cytometry for Immune Cell Profiling

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BM cells were incubated with Fc Block (BD Bioscience) for 15 min, and then stained with the following conjugated antibodies (30 min at 4°C in the dark): CD45-APC-Cy7, NK1.1-APC, CD3-APC, CD19-APC, CD11b-PerCP-Cy5.5, Ly6C-FITC (all from BD Bioscience), TER119-APC, Ly6G-eFluor450 (eBioscience) for neutrophil and monocyte populations, CD45-APC-Cy7, CD19-PE, CD4-V500, CD8-PerCP-Cy5.5, Gr1-APC (BD Bioscience), and CD3-eFlour450, NK1.1-PE-Cy7, TER119-APC (eBioscience) for lymphocyte population. After antibody staining, the cells were incubated with 7-AAD for 5 min at room temperature as a viability dye for dead cell exclusion and analyzed in the presence of AccuCheck counting beads by FACSSymphony A3. The data were further analyzed by using the FlowJo software.
Kim Y., Lee Y., Lee M.N., Nah J., Yun N., Wu D, & Pae M. (2022). Time-restricted feeding reduces monocyte production by controlling hematopoietic stem and progenitor cells in the bone marrow during obesity. Frontiers in Immunology, 13, 1054875.
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3

Human Melanoma Cell Isolation and Sorting

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All procedures have been described before (23 (link)). All antibody labeling was performed on ice for 20 min, followed by washing and centrifugation. Cells were stained with directly conjugated antibodies to mouse CD45 (30-F11-APC, eBioscience), mouse CD31 (390-APC, BioLegend), Ter119 (TER-119-APC, eBioscience), and human HLA-A, B, C (G46-2.6-BV421, BD Biosciences) to select live human melanoma cells and exclude endothelial and hematopoietic cells. Cells were examined on an LSRFortessa cell analyzer (BD Biosciences) or sorted on a FACSAria cell sorter (Becton Dickinson). SETDB1 or EZH2 knockdown cells were sorted as DsRed and green fluorescent protein (GFP) double-positive cells.
Shi X., Tasdogan A., Huang F., Hu Z., Morrison S.J, & DeBerardinis R.J. (2017). The abundance of metabolites related to protein methylation correlates with the metastatic capacity of human melanoma xenografts. Science Advances, 3(11), eaao5268.
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4

Multicolor Flow Cytometry Immunophenotyping

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Cells were stained for 30 minutes on ice with antibodies against CD11b PE-Cy7 (Biolegend), Gr1 PE (eBioscience, 1:400), CD45R/B220 FITC (BD), CD3ε APC-Cy7 (Biolegend), and Ter119 APC (eBioscience) and analyzed using a LSRII flow cytometer (BD).
Yu W., Goncalves K.A., Kishikawa H., Li S., Sun G., Yang H., Vanli N., Wu Y., Jiang Y., Hu M.G., Friedel R.H, & Hu G.F. (2017). Plexin-B2 mediates physiologic and pathologic functions of angiogenin. Cell, 171(4), 849-864.e25.
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5

Antibody Panel for Cellular Proteins

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The antibodies against the following proteins were used as follows: EPB41 (rabbit, Sigma; 22, 23), Giα3 (rabbit, Millipore), NuMA (rabbit, Cell Signaling Technology; rabbit, Abcam), LGN (rabbit, EMD Millipore Corp; Dr Quansheng Du, Georgia Regents University), p150Glued (mouse, BD Transduction Laboratories), dynein (IC 74.1, Chemicon), Numb (mouse, Millipore; rabbit, Abcam; Rabbit, Novus Biologicals), Nestin (mouse, Developmental Studies Hybridoma Bank), α-Hemoglobin (rabbit, Santa Cruz biotechnology), Notch1 (mouse, BioLegend), Hes1 (mouse, OriGene Technologies), pericentrin (rabbit, Covance), β-actin (rabbit, GeneTex; AC-74, Sigma-Aldrich), γ-tubulin (T5192, rabbit, Millipore Sigma; clone GTU88, Sigma-Aldrich), α-tubulin (mouse, Calbiochem; mouse, Sigma; clone YL1/2; EMD Millipore), GAPDH (mouse, Sigma), mCherry (Rabbit, BioVision), Flag (mouse, Sigma), Ter-119 APC (Rat, eBioscience), CD71 FITC (Rat, BD Pharmingen).
Huang S.C., Vu L.V., Yu F.H., Nguyen D.T, & Benz EJ J.r. (2021). Multifunctional protein 4.1R regulates the asymmetric segregation of Numb during terminal erythroid maturation. The Journal of Biological Chemistry, 297(3), 101051.
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6

Multicolor Flow Cytometry Panel

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Antibodies used in this study included CDK6 (C-21) from Santa Cruz), α-Tubulin from Sigma, RUNX1 from Abcam, and Sca-1-FITC/APC, CD36-APC, CD31-APC, CD45-APC, TER119-APC, CD24-FITC, CD29-FITC, CD34-FITC from e-Bioscience, BrdU monoclonal antibody (MA3-071) from ThermoFisher, eBioscience™ BrdU Staining Kit for Flow Cytometry FITC from ThermoFisher (8811-6600-42), and 7-Aminoactinomycin D (7-AAD) from ThermoFisher (A1310).
Hu A.J., Li W., Pathak A., Hu G.F., Hou X., Farmer S.R, & Hu M.G. (2023). CDK6 is essential for mesenchymal stem cell proliferation and adipocyte differentiation. Frontiers in Molecular Biosciences, 10, 1146047.
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7

Isolation and Analysis of Mesenchymal Progenitor Cells

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Freshly isolated mPDC, mBMC-F and mBMC-B were resuspended in PBS supplemented with 2% fetal bovine serum (FBS; Gibco) and 2 mM EDTA. The cells were incubated for 30 min at 4 °C with the respective antibodies at the indicated concentration (Table 1). Flow cytometry analysis was performed on a Gallios Flow Cytometer (Beckman Coulter, Brea, USA) and analyzed by using the Kaluza software (Beckman Coulter). Due to the high amount of hematopoietic cells in the bone marrow fraction, a Lineage Cell Depletion Kit and CD45 MicroBeads were used together with LD columns (all from Miltenyi biotech, Bergisch Gladbach, Germany) according to manufacturer's instructions before investigating the panels of markers described in Chan et al. [12] (link).
To obtain Nes-GFP + cells from the periosteum, mPDC from Nes-GFP + mice were isolated and incubated with Ter119-APC, CD45-APC and CD31-APC antibodies (eBioscience; Thermo Fisher Scientific, Asse, Belgium). The Nes-GFP -and Nes-GFP + cells in the Ter119 -CD45 -CD31 -population were then sorted by flow cytometry (BD FACSAria III, BD Biosciences, Erembodegem, Belgium).
Tournaire G., Stegen S., Giacomini G., Stockmans I., Moermans K., Carmeliet G, & van Gastel N. (2020). Nestin-GFP transgene labels skeletal progenitors in the periosteum. Bone, 133.
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8

Quantification of Cell Proliferation and Apoptosis

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Assessment of cell proliferation was performed by administering 5-ethynyl-2ʹ-deoxyuridine (EdU) (Life Technologies Corporation, A10044) intraperitoneally to adult mice. Each mouse received a total of 1.5 mg of EdU dissolved in PBS in three divided doses at 2-hour intervals over 6-hours and then sacrificed. Cells were isolated as described above in the flow cytometry section. Evaluation of EdU-positive cells was performed using Click-iT Imaging Kit with Alexa Fluor 488 Assay Kit (Life Technologies Corporation, Carlsbad, CA, USA; C10632). To quantify apoptotic cells, tibias and femurs were harvested from adult mice and stained with 1:250 anti-CD45-APC (eBioscience, San Diego, CA, USA; 17–0451–82) and Ter119-APC (eBioscience, 17–5921–82) followed by staining with FITC Annexin V Apoptosis Detection Kit (BioLegend, San Diego, CA, USA; 640922) for 15 minutes. The rate of apoptosis was measured and evaluated under flow cytometry using an LSR II flow cytometer.
Balani D.H., Trinh S., Xu M, & Kronenberg H.M. (2021). Sclerostin Antibody Administration Increases the Numbers of Sox9creER+ Skeletal Precursors and Their Progeny. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 36(4), 757-767.
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9

Isolation and Characterization of Murine Mammary Cells

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Mammary glands were dissected from 10- to 12-week-old female mice at pregnancy day 14.5, cut into pieces, and placed in DMEM/F12 (Gibco) with 5% FBS (Gibco), 300 U/mL collagenase (Gibco) and 100 U/mL hyaluronidase (Sigma) for 2 h at 37°C. After digestion, red blood cells were removed with NH4Cl (STEMCELL Technologies, Vancouver, BC, Canada), and then a single-cell suspension was obtained by sequential dissociation in 37°C preheated 0.25% trypsin (Gibco) for 2 min and in 5 mg/mL Dispase (STEMCELL Technologies) containing 0.1 mg/mL DNase I (Sigma) for 5 min with pipetting, followed by filtration through a 40 μm filter (BD, San Jose, CA, United States). The single-cell suspension was stained with specific antibodies in DPBS containing 2% FBS for 15 min on ice. After being washed three times with DPBS containing 2% FBS, cells were analyzed by FACS Verse or sorted by FACS Aria. The following antibodies were used: CD45-APC (17-0451-82, eBioscience, San Diego, CA, United States), CD31-APC (17-0311-82, eBioscience), TER119-APC (17-5921-82, eBioscience), CD24-PE-Cy7 (25-0242-82, eBioscience), CD29-FITC (11-0291-82, eBioscience), CD61-PE (561910, BD), and Fixable Viability Dye eFluor 450 (65-0863-14, eBioscience). Data analysis was performed with FlowJo 7.6.1.
Shao C., Lou P., Liu R., Bi X., Li G., Yang X., Sheng X., Xu J., Lv C, & Yu Z. (2021). Hormone-Responsive BMP Signaling Expands Myoepithelial Cell Lineages and Prevents Alveolar Precocity in Mammary Gland. Frontiers in Cell and Developmental Biology, 9, 691050.
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10

Comprehensive Flow Cytometry Profiling of Bone Marrow Stroma

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For flow cytometry and FACS, cells were resuspended in Media 199 supplemented with 2% FBS and stained for Ter119-APC (eBioscience, Ref#17–5921–82, clone TER–119), CD71-PECy7 (Biolegend, Ref#113812, clone RI7217), CD45-PE (eBioscience, Ref#12–0451–82, clone 30–F11), CD3-PE (Biolegend, Ref#100206, clone 17A2), B220-PE (Biolegend, Ref#103208, clone RA3–6B2), CD19-PE (Biolegend, Ref#115508, clone 6D5), Gr-1-PE (Biolegend, Ref#108408, clone RB6–8C5), and Cd11b-PE (Biolegend, Ref#101208, clone M1/70) for 30 minutes on ice. Dead cells and debris were excluded by FSC, SSC, 7-AAD (ThermoFisher Scientific, Ref#00–6993–50) and Calcein AM (ThermoFisher Scientific, Ref#C3099) profiles. FACS and cytometry was performed on a BD FACSAria II sorter, and sorted bone marrow stromal cells were collected in Media 199 supplemented with 2% FBS and 0.4% UltraPure BSA (ThermoFisher Scientific, Ref#AM2616). Bone marrow stroma was enriched by sorting of live cells (7-AAD-/Calcein+) negative for erythroid (CD71/Ter119) and immune lineage markers (CD45/CD3/B220/CD19/Gr-1/CD11b). For flow cytometric analysis (e.g. Figure S1K), we used Ly6a/Sca-1 (Biolegend, Ref#108127, clone), CD31 (Pecam1) (BD Bioscience, Ref#565097), Vcam1 (Biolegend, Ref#105710), CD34 (eBioscience, Ref#11–0341–82), Cdh5/CD144 (Biolegend, Ref#138006)
Baryawno N., Przybylski D., Kowalczyk M.S., Kfoury Y., Severe N., Gustafsson K., Kokkaliaris K.D., Mercier F., Tabaka M., Hofree M., Dionne D., Papazian A., Lee D., Ashenberg O., Subramanian A., Vaishnav E.D., Rozenblatt-Rosen O., Regev A, & Scadden D.T. (2019). A cellular taxonomy of the bone marrow stroma in homeostasis and leukemia. Cell, 177(7), 1915-1932.e16.
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