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Ter119

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Ter119 is a monoclonal antibody used for the identification and isolation of erythroid cells. It recognizes the Ter119 antigen, which is expressed on the surface of red blood cells and their precursors. Ter119 is a widely used marker for the analysis and purification of erythroid lineage cells in flow cytometry and cell sorting applications.

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Lab products found in correlation

Biomag goat anti rat igg beads, by Qiagen (2 mentions) Corn oil control, by Merck Group (1 mentions) Anti rat igg coupled magnetic beads, by Qiagen (1 mentions) 4 hydroxytamoxifen oht, by Merck Group (1 mentions) N acetyl cysteine (nac), by Merck Group (1 mentions) Rapamycin, by LC Laboratories (1 mentions) Metformin, by Merck Group (1 mentions) Indo 1 am, by Merck Group (1 mentions) Low melt agarose, by Lonza (1 mentions) Vt1000s vibratome, by Leica (1 mentions) Ter119, by BioLegend (1 mentions) Mr9 4, by BioLegend (1 mentions) Pc 61, by BioXCell (1 mentions) M1 70, by BioXCell (1 mentions) Ebio927, by Thermo Fisher Scientific (1 mentions) M1 70, by BioLegend (1 mentions) Rb6 8c5, by BioXCell (1 mentions)

5 protocols using ter119

1

Adoptive NK Cell Transfer and Modulation

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T, B, and red blood cells were labeled with 10 μg per spleen of rat monoclonal antibodies against CD4 (GK1.5), CD8 (53.6.72), CD19 (1D3), and Ter119 (Bio-X-cell) and magnetically depleted from total splenocyte suspensions with the use of anti-rat IgG-coupled magnetic beads (QIAGEN). In all experiments, approximately 2 × 105 enriched NK cells were injected intravenously into mice. In adoptive co-transfer experiments, equal numbers of Ly49H+ NK cells from each population (CD45.1+ and CD45.2+) were injected into recipients 1 day before infection. In some experiments, recipient mice were injected intraperitoneally with 1 mg/day of hydroxytamoxifen (4-OHT) dissolved in corn oil or corn oil control (Sigma) for 5 consecutive days starting on day 4 after infection and subsequently injected intraperitoneally with PBS, 200 μg/day metformin (Sigma), 600 μg/kg/day rapamycin (LC Laboratories), or 1.25 mg/day NAC (Sigma) from day 8 to 28 after infection.
O’Sullivan T.E., Johnson L.R., Kang H.H, & Sun J.C. (2015). BNIP3- and BNIP3L-Mediated Mitophagy Promotes the Generation of Natural Killer Cell Memory. Immunity, 43(2), 331-342.
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2

Isolation of Immune Cells from Tissues

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Spleens and liver were dissociated through a 100-μm strainer. Dissociated liver was resuspended in a 40% Percoll solution and centrifuged at 700 × g (with reduced break speed) for 12 min to separate lymphocytes from hepatocytes. To isolate bone marrow cells, femur and tibia were ground with mortar and pestle and the resulting solution was filtered through a 100-μm strainer. Red blood cells in spleen, liver, blood, and bone marrow were lysed using ACK lysis buffer. In experiments involving sorted cells, splenocytes were incubated with rat anti-mouse CD3ε and CD8α (except when CD8+ T cells were required) along with Ly6G, CD4, CD19, and Ter-119 antibodies (Bio X Cell, clones 17A2, 2.43, 1A8, GK1.5, 1D3, and Ter-119, respectively) followed by incubation with BioMag goat anti-rat IgG beads (QIAGEN) to remove Ab-bound cells without touching populations of interest.
Sheppard S., Santosa E.K., Lau C.M., Violante S., Giovanelli P., Kim H., Cross J.R., Li M.O, & Sun J.C. (2021). Lactate dehydrogenase A-dependent aerobic glycolysis promotes natural killer cell anti-viral and anti-tumor function. Cell reports, 35(9), 109210.
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3

Thymocyte Motility Imaging by Two-Photon Microscopy

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CD4SP thymocytes were enriched by magnetic depletion using antibodies against CD8, Gr-1, Ter119, B220, CD25, and CD11b (BioXCell). 1×106Ebi2+/+ and Ebi2-/- CD4SP cells were stained with Indo-1AM (Sigma) and CMTPX red (Life Technologies), respectively, for 30min at 37°C,according to manufacturers’ instructions and mixed at a 1:1 ratio in complete RPMI. 3-4 week old pCX-EGFP thymi were embedded in low-melt agarose (Lonza) and vibratome sectioned, with a VT1000S Vibratome (Leica), as previously described [12 (link)]. After incubating thymocytes on slices for 1-2 h at 37°C 5% CO2, two-photon images were acquired every 15 s, through a depth of 40 μm at 5-μm intervals using an Ultima IV microscope equipped with a 20x NA 1.0 water-immersion objective, and PraireView software (Prairie). MaiTai lasers (SpectraPhysics) tuned to 740 nm and 900 nm were used to excite Indo-1 and EGFP/CMTPX, and emitted light was passed through 400/50, 480/40, 535/50, and 607/45 filters (Chroma Technology) for detection of the two Indo1 emission peaks, EGFP, and CMTPX, respectively. were carried out with Imaris v.8 (Bitplane).
Ki S., Thyagarajan H.M., Hu Z., Lancaster J.N, & Ehrlich L.I. (2017). EBI2 contributes to the induction of thymic central tolerance by promoting rapid motility of medullary thymocytes. European journal of immunology, 47(11), 1906-1917.
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4

Characterizing Mouse Immune Cell Phenotypes

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Antibodies directed against the following mouse markers were used: CD3 (145–2C11, Tonbo Biosciences 60–0031), CD4 (RM4–5, BioLegend 100559; GK1.5, BioXCell BE0003–1), CD8 (53–6.7, Tonbo Biosciences 75–0081; 53.6.72, BioXCell BE0004), CD11b (M1/70, BioLegend 101204; M1/70, BioXCell BE0007), CD11c (N418, BioLegend 117322), CD19 (6D5, BioLegend 115534), CD25 (PC61, BioLegend 102024; PC61.5.3, BioXCell BE0012), CD45 (30-F11, BioLegend 103138), CD45.1 (A20, BioLegend 110714), CD69 (H1.2F3, BioLegend 104504), CD80 (16–10A1, BioLegend 104724), B220 (RA3–6B2, BioLegend 103232; RA3.3A1/6.1, BioXCell BE0067), AIRE (5H12, eBioscience 53–5934–82), EpCAM (G8.8, BioLegend 118206), F4/80 (BM8, BioLegend 123116), Gr-1 (RB6–8C5, BioLegend 108410; RB6–8C5, BioXCell BE0075), I-A/I-E (M5/114.15.2, BioLegend 107628), NK1.1 (PK136, BioLegend 108716), PDCA1 (eBio927, eBioScience 12–3172–82), Sirpα (P84, BioLegend 144008), TER-119 (TER-119, BioLegend 116210; TER-119, BioXCell BE0183), Vα2 (B20.1, BioLegend 127818), Vβ5 (MR9–4, BioLegend 139504), and XCR1 (ZET, BioLegend 148212). For immunostaining ~107 cells in 100 μL of PBS + 2% bovine calf serum (BCS), fluorochrome-conjugated antibodies were diluted from stock concentrations of 0.5 mg mL−1 and incubated with cells for 20 min on ice, unless specified.
Lancaster J.N., Thyagarajan H.M., Srinivasan J., Li Y., Hu Z, & Ehrlich L.I. (2019). Live-cell imaging reveals the relative contributions of antigen-presenting cell subsets to thymic central tolerance. Nature Communications, 10, 2220.
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5

Isolation of Immune Cells from Tissues

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Spleens and liver were dissociated through a 100-μm strainer. Dissociated liver was resuspended in a 40% Percoll solution and centrifuged at 700 × g (with reduced break speed) for 12 min to separate lymphocytes from hepatocytes. To isolate bone marrow cells, femur and tibia were ground with mortar and pestle and the resulting solution was filtered through a 100-μm strainer. Red blood cells in spleen, liver, blood, and bone marrow were lysed using ACK lysis buffer. In experiments involving sorted cells, splenocytes were incubated with rat anti-mouse CD3ε and CD8α (except when CD8+ T cells were required) along with Ly6G, CD4, CD19, and Ter-119 antibodies (Bio X Cell, clones 17A2, 2.43, 1A8, GK1.5, 1D3, and Ter-119, respectively) followed by incubation with BioMag goat anti-rat IgG beads (QIAGEN) to remove Ab-bound cells without touching populations of interest.
Sheppard S., Santosa E.K., Lau C.M., Violante S., Giovanelli P., Kim H., Cross J.R., Li M.O, & Sun J.C. (2021). Lactate dehydrogenase A-dependent aerobic glycolysis promotes natural killer cell anti-viral and anti-tumor function. Cell reports, 35(9), 109210.
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