Ter119 biotin

Manufactured by Thermo Fisher Scientific

Sourced in United States

Ter119-Biotin is a cell surface marker that binds to the Ter119 antigen, which is expressed on erythroid cells. It is a useful tool for the identification and isolation of erythroid cells in flow cytometry and cell sorting applications.

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Ter119 (107 citations) Ter119-biotin (clone Ter119, (2 citations) Anti-Ter119-biotin (2 citations) Biotin-conjugated anti-Ter119 antibody (2 citations)

5 protocols using ter119 biotin

Fetal Blood Cell Staining and Imaging

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Fetal blood samples were stained with Hoechst 33342 (Molecular Probes, Eugene, OR) and MitoTracker Deep Red (Invitrogen) according to manufacturer directions. Cells were then blocked and stained in 5% rat whole serum (Invitrogen) prepared in PB2 (Dulbecco PBS and 0.1% glucose [Invitrogen] and 0.3% BSA [Gemini BioProducts]) with 1:200 dilutions of PE-CD71, Biotin-Ter119 (eBioscience, San Diego, CA), and 2 μg/mL of thiazole orange (Sigma-Aldrich). After washing, cells were stained with 1:250 PE-Texas Red streptavidin (BD Biosciences, San Jose, CA). Data were collected on the ImageStreamX and analyzed using IDEAS 6.2 (Luminex, Austin, TX); debris, calibration beads, and cell aggregates were excluded by gating on size and aspect ratios, and focused cells were selected based on sharpness of the brightfield stain (gradient root mean square) as previously described.^{25 (link)}

Nemkov T., Kingsley P.D., Dzieciatkowska M., Malik J., McGrath K.E., Hansen K.C., D’Alessandro A, & Palis J. (2022). Circulating primitive murine erythroblasts undergo complex proteomic and metabolomic changes during terminal maturation. Blood Advances, 6(10), 3072-3089.

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Comprehensive Multiparameter Flow Cytometry Protocol

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Single‐cell suspensions from the spleen and bone marrow were counted using counting slides (SD‐100, Nexcelom) in a Cellometer Mini Automated Cell Counter (Nexcelom). Surface antigens were stained with demonstrated conjugated primary antibodies in the staining buffer (1× PBS, 3% BSA, 1 mM EDTA, 0.1% NaN3) at 4 °C for 30 min. Antibodies used were as follows: APC TER‐119 (557 909, BD), PE CD71 (553 267,BD), Biotin CD3 (100 244, Biolegend), Biotin CD45R (13‐0452‐81, eBioscience),Biotin CD4 (13‐0041‐81, eBioscience), Biotin CD5 (13‐0051‐81, eBioscience), Biotin CD8a Biotin (13‐0081‐81, eBioscience),Biotin CD19 (13‐0193‐81, eBioscience),Biotin CD11b (13‐0112‐81, eBioscience),Biotin Ly‐6G (13‐5931‐81, eBioscience), Biotin TER‐119 (13‐5921‐81, eBioscience), PE‐Cy7 Streptavidin (25‐4317‐82, eBioscience), APC Ly‐6A/E (Sca‐1) (17‐5981‐82, eBioscience), APC‐EFLUOR CD117 (C‐KIT) (47‐1172‐82, eBioscience). After staining, cells were washed once with 1× PBS and quickly analyzed by a cytoflex S flow cytometer (cytoflex S, Beckman Coulter). All analyses were performed using CytExpert software (CytExpert, Beckman Coulter, Inc.).
An auto hematology analyzer (BC‐2800Vet, Mindray) was used to analyze diluted peripheral blood that had been collected in an anticoagulation tube and diluted in EDTA buffer (0.5 M EDTA, pH 8.0) at a quantitative ratio of 1:1.

Wu X., Wang Y., Chen B., Liu Y., Li F., Ou Y., Zhang H., Wu X., Li X., Wang L., Rong W., Liu J., Xing M., Zhao X., Liu H., Ge L., Lv A., Wang L., Wang Z., Li M, & Zhang H. (2023). ABIN1 (Q478) is Required to Prevent Hematopoietic Deficiencies through Regulating Type I IFNs Expression. Advanced Science, 11(3), 2303555.

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Multiparametric Flow Cytometric Analysis

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Cell suspensions of different organs were lysed with ammonium chloride buffer (0.150 mM NH₄Cl, 0.1 mM EDTA, 0.150 mM KHCO₃) to eliminate erythrocytes followed by staining with CD19-FITC/APC (eBio1D3, 1:100), CD38-PE (clone 90, 1:100), IgM-PE/PECy7 (II/41, 1:200), IgD-eFlour450 (11-26c, 1:30), CD5-APC/Pacific Blue (53-7.3, 1:80), B220-eFlour605 (RA3-6B2, 1:40), CD3-FITC (145-2C11, 1:100), ZAP70-FITC (1E7.2, 1:20), CD21-FITC (eBio8D9, 1:100), CD23-PECy7 (B3B4, 1:160), CD93-PE (AA4.1, 1:80), LIN-Cocktail (CD11b-Biotin (M1/70, 1:130), CD3-Biotin (145-2C11, 1:130), Ter119-Biotin (Ter119, 1:130) and Gr1-Biotin (RB6-8C5, 1:130), Streptavidin-PerCP (1:160), Annexin V-Pacific Blue (1:25) and 7-AAD (1:100) (all from eBioscience). Cells were analysed with the Canto II cytometer (BD Bioscience). Data was obtained with the BD FACSDiva^TM (BD) and analysed with the FlowJo 8.5.3 software. The sequential gating strategy for all FACS panels is provided in Supplementary Fig. 11.

Märklin M., Heitmann J.S., Fuchs A.R., Truckenmüller F.M., Gutknecht M., Bugl S., Saur S.J., Lazarus J., Kohlhofer U., Quintanilla-Martinez L., Rammensee H.G., Salih H.R., Kopp H.G., Haap M., Kirschniak A., Kanz L., Rao A., Wirths S, & Müller M.R. (2017). NFAT2 is a critical regulator of the anergic phenotype in chronic lymphocytic leukaemia. Nature Communications, 8, 755.

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Comprehensive Immune Cell Profiling

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All antibodies are from BD Biosciences, except where noted otherwise. CD3-biotin, CD11b-FITC -PE -PE-Cy7 -biotin, CD16/32-PE, CD19-PE, CD24-PE-Cy7, CD34-FITC, CD43-PE, CD48-FITC, CD95-PE, CD150-PE-Cy7, c-Kit-APC -FITC (eBioscience), Sca-1-PE – PE-Cy7 (eBioscience), B220-biotin, FLT3-PE (eBiosciences), Gr-1-FITC -biotin, IgM-APC, IgD-FITC (eBioscience), IL-7R-APC -biotin, TER119-biotin, GL7-FITC and TCR-APC. Cells incubated with biotinylated antibodies were treated with streptavidin conjugated with APC-Cy7 (BD Biosciences). Multipotent progenitors (MPPs) as FLT3⁺ LSK cells, common lymphoid progenitors (CLPs) as Lin⁻, IL-7R⁺ Sca-1^lo c-Kit^lo; granulocyte macrophage progenitors (GMPs) as Lin⁻, CD16/32^hi, CD34⁺, Sca1⁻, c-Kit⁺; common myeloid progenitors (CMPs) as Lin⁻, CD16/32^lo, CD34⁺, Sca1⁻, c-Kit⁺; and megakaryocyte erythroid progenitors (MEPs) as Lin⁻_, CD34⁻, CD16/32⁻, Sca-1⁻,c-Kit⁺. Pre-B, pro-B and prepro-B cells were determined by CD24 and CD43 staining of B220⁺, IgM⁻ bone marrow cells.

Duyvestyn J.M., Taylor S.J., Dagger S.A., Orandle M., Morse HC I.I.I., Thien C.B, & Langdon W.Y. (2014). Dasatinib Targets B-Lineage Cells but Does Not Provide an Effective Therapy for Myeloproliferative Disease in c-Cbl RING Finger Mutant Mice. PLoS ONE, 9(4), e94717.

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Flow Cytometry Antibody Staining Protocol

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For the flow cytometry analyses, cells were stained with monoclonal antibodies against the following anti-mouse molecules: CD3-biotin, Ter119-biotin, GR-1-biotin, B220-biotin, CD11b-biotin, NK1.1-biotin, CD3-biotin, CD4-biotin, CD8-biotin, cKit-BV650, Sca1-PE, CD135-PerCP, CD34-APC, CD25-PE, CD44-APC, CD3-APC, CD8-PerCP, CD-BV650, CD45.1/Ly5.1-PE-Cy7, CD45.2/Ly5.2-APC-Cy7 (all from eBiosciences, CA, USA). For secondary detection of biotinylated antibodies, streptavidin conjugated with FITC or PE-Cy7 was used (eBiosciences, San Diego, CA, USA). All flow cytometry measurements (Canto II BD biosciences or LSRII BS Biosciences) were calibrated first with BD™ CompBead Plus, κ/Negative Control (BSA) Compensation Plus (7.5 µm). Recommended flow cytometry settings are specified in [17 ]. Flow cytometry analyses were performed using FlowJo software (Treestar, Ashland, OR, USA).

de Roo J.J., Chhatta A., Garcia-Perez L., Naber B.A., Vloemans S.A., Salvatori D.C., Pike-Overzet K., Mikkers H, & Staal F.J. (2022). Axin2/Conductin Is Required for Normal Haematopoiesis and T Lymphopoiesis. Cells, 11(17), 2679.

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