Ter119 pe

Manufactured by BD

Ter119-PE is a fluorescent-conjugated antibody used for flow cytometry analysis. It targets the Ter119 antigen, which is expressed on mouse erythroid cells. Ter119-PE can be used to identify and quantify erythroid cells in mouse samples.

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Lab products found in correlation

Facscalibur, by BD (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Alexa fluor 647, by Abcam (1 mentions) Csf1r, by Abcam (1 mentions) Runx1, by Abcam (1 mentions) C kit fitc, by Thermo Fisher Scientific (1 mentions) Alexa fluor tagged secondary antibodies, by Thermo Fisher Scientific (1 mentions) Cd34 fitc, by Thermo Fisher Scientific (1 mentions) Ve cadherin, by R&D Systems (1 mentions) Facs lsr 2 instrument, by BD (1 mentions) Click it edu alexa fluor 647 imaging kit, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Corning (1 mentions) Nk1.1 pe, by BD (1 mentions) B220 pe, by BD (1 mentions) Biotin conjugated, by BD (1 mentions) Streptavidin pacific blue, by Thermo Fisher Scientific (1 mentions) C kit apc, by BD (1 mentions) Cd4 pe, by BD (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions) Lsrfortessa, by BD (1 mentions)

7 protocols using ter119 pe

Multiparametric Flow Cytometry and IHC Assays

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The following antibodies were used for flow cytometry and immunohistochemistry analysis: CD31 (BD Bioscience, #553371; DSHB, 2H8), CD34-FITC (eBiosciences, #11-0341), Ter119-PE (BD Biosciences, #553673), Runx1 (Abcam, ab92336), c-kit-FITC (eBiosciences, #11-1171), CSF1R (Abcam, #Ab32633; eBioscience AFS98), and VE-Cadherin (R&D, #AF1002). Alexa-Fluor tagged secondary antibodies (Invitrogen) were used for all non-conjugated primary antibodies with the exception of anti-Armenian Hamster secondary (AlexaFluor647, Abcam).

Kasaai B., Caolo V., Peacock H.M., Lehoux S., Gomez-Perdiguero E., Luttun A, & Jones E.A. (2017). Erythro-myeloid progenitors can differentiate from endothelial cells and modulate embryonic vascular remodeling. Scientific Reports, 7, 43817.

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Erythroid Cell Isolation and Analysis

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The cells from BM, spleen, and fetal liver were passed through a 23-G needle and filtered through a 70-μM cell strainer to obtain single-cell suspensions. The cells were stained with antibodies, including CD71 (FITC), TER119 (PE), CD44 (APC), CD45 (PE-Cy7), Gr1 (PE-Cy7), CD11b (PE-Cy7). All of the antibodies were purchased from BD Pharmingen. Following incubation of cells with conjugated antibodies for 30 minutes at 4 °C, cells were washed in PBS containing 0.5% bovine serum albumin (BSA) and analyzed using a FACS Calibur cell analyzer (BD Biosciences). For enucleation analysis, BM cells from Asxl1^f/f and Asxl1^∆/∆ mice were stained with TER119-PE (BD Pharmingen) for 30 mins at room temperature and nuclear staining was performed with 10 μg/mL Hoechst 33342. Data were acquired on FACS LSR II instrument (BD Biosciences).

Shi H., Yamamoto S., Sheng M., Bai J., Zhang P., Chen R., Chen S., Shi L., Abdel-Wahab O., Xu M., Zhou Y, & Yang F.C. (2016). ASXL1 plays an important role in erythropoiesis. Scientific Reports, 6, 28789.

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Flow Cytometry Analysis of Immune Cells

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For all flow cytometry analysis cells were stained in PBS (Corning Cellgro) supplemented with 2% of inactivated FBS (Gemini BioProducts). The following antibodies were used: B220 PE, CD11b PE or APC, CD11C PE or APC, CD4 PE, CD8 PE, NK1.1 PE, Ter119 PE, CD3 PE, c-Kit APC or biotin-conjugated (BD Biosciences). Sca-1 PEcy7, streptavidin-APC (eBiosciences) and streptavidin Pacific blue (Invitrogen). DAPI was used to exclude dead cells. Edu incorporation assays were performed according to manufacturer’s protocol (Invitrogen, Click-iT Edu Alexa Fluor 647 Imaging kit), with cells pulsed with Edu for 30 minutes. Cells were co-stained with DAPI for DNA content measurement. Annexin V apoptosis staining was performed according to manufacturer’s protocol (BD Pharmingen, APC Annexin V). All flow cytometry was performed on a LSR Fortessa (BD Biosciences).

Kaushik S., Liu F., Veazey K., Gao G., Das P., Neves L., Li K., Zhong Y., Lu Y., Giuliani V., Bedford M.T., Nimer S.D, & Santos M.A. (2017). Genetic deletion or small molecule inhibition of the arginine methyltransferase PRMT5 exhibit anti-tumoral activity in mouse models of MLL-rearranged AML. Leukemia, 32(2), 499-509.

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Multiparameter Flow Cytometry Analysis

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CD23 (B3B4)-PE and -BV421[1:200], TCRβ (H57-597)-PE[1:400], Ter-119-PE[1:400], CD138 (281-2)-PE[1:400] and XBP1s (Q3-695)-PE-CF594[1:200] were purchased from BD Biosciences. IgD (11-26.2a)-APC-Cy7[1:200], CD4 (GK1.5)-PE-Cy7[1:400], Ter-119-PE-Cy7[1:400], F4-80 (BM8)-PE-Cy7[1:400], Sca-1 (D7)-BV605[1:200], B220 (RA3-6B2)-BV421[1:200], and CD138 (281-2)-BV605[1:400] were purchased from BioLegend. ATF-4 (D4B8)[1:1000], XBP1s (E8Y5F) [1:1000], p58IPK (C56E7)[1:1000], ATF-6 (D4Z8V) [1:1000], BiP (C50B12)-PE[1:200], pAKT (S473) (D9E)-PE[1:100] and pS6(S235/236) (D57.2.2E)-PE-Cy7[1:200] were purchased from Cell Signaling Technology. AA4.1-APC[1:100], CD21/35 (8D9)-PE-Cy7[1:200], IgM (11/41)-PerCP-ef710[1:200], and CD8α (53-6.7)-PE and -PE-Cy7[1:400] were purchased from eBioscience (ThermoFisher). CD19 (6D5)-APC-Cy5.5[1:150] and F4-80 (BM8)-PE[1:400] were purchased from Invitrogen. B220 (RA3-6B2)-APC[1:200] was purchased from Tonbo biosciences.

Gaudette B.T., Jones D.D., Bortnick A., Argon Y, & Allman D. (2020). mTORC1 coordinates an immediate unfolded protein response-related transcriptome in activated B cells preceding antibody secretion. Nature Communications, 11, 723.

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Red Blood Cell Analysis in Mice

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Blood was collected from either cardiac puncture post-euthanasia or via tail vein. Red cell indices were measured using the Advia 2120 (Siemens) or Hemavet (Dres Scientific) automated blood analyser. For reticulocyte analysis on peripheral red blood cells, 1l aliquots of blood were incubated in 0.5mL thiazole orange (100ng/mL, Sigma) and 0.2mL Ter119-PE (BD Pharmingen, catalogue number 553673) for 30 min at room temperature protected from light then analyzed using a FACSCalibur flow cytometer (BD Biosciences).
Serum EPO was quantified using the R&D Systems Quantikine ELISA kit (Catalogue number MEP00B) according to manufacturer's instructions.

McManus J.F., Nguyen N.N., Davey R.A., MacLean H.E., Pomilio G., McCormack M.P., Chiu W.S., Wei A.H., Zajac J.D, & Curtis D.J. (2020). Androgens stimulate erythropoiesis through the DNA-binding activity of the androgen receptor in non-hematopoietic cells. European journal of haematology, 105(3).

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Erythroblast Isolation and Analysis

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For analyzing mice erythroblasts, BM cells from P38α^+/− and P38α^−/− mice were collected and washed with staining buffer (PBS + 5% FBS). Then cells were subsequently stained for 30 min on ice with the following anti-mouse antibodies at a 1:50 dilution:CD71-FITC (553266), Ter119-APC (557909) or Ter119-PE (553673) from BD Biosciences. To evaluate human erythroid differentiation, collected and washed cells were incubated for 30 min on ice with the following anti-human antibodies at a 1:50 dilution: CD235a-PE (12-9987-82, eBioscience), CD71-PerCP/Cy5.5 (334114,Biolegend) or CD71-APC (334108, Biolegend). After wash with staining buffer, the cell pellets were resuspended in staining buffer supplemented with DAPI (Thermo Fisher Scientific) or Propidium iodide (PI) (Sigma-Aldrich). All samples were measured with a BD LSR-II or FACS Calibur (BD Biosciences) and analyzed with Flowjo (Treestar, Ashland, OR, USA).

Hu P., Nebreda A.R., Hanenberg H., Kinnebrew G.H., Ivan M., Yoder M.C., Filippi M.D., Broxmeyer H.E, & Kapur R. (2018). P38α/JNK signaling restrains erythropoiesis by suppressing Ezh2-mediated epigenetic silencing of Bim. Nature Communications, 9, 3518.

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Bone Marrow Cell Isolation and Transplantation

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PanRosa^YFP mice were used as donors. Bone marrow cells of PanRosa^YFP mice were flushed from femurs, tibias, coxae, and humeri using PBS supplemented with 5% heat-inactivated FCS. Cells were filtered through a 40-µm cell strainer (Falcon). Fc receptors were blocked by incubating cells in 5% FCS with purified mouse IgG (500 mg/ml; Jackson ImmunoResearch). All stainings were performed in 5% FCS on ice for 30 min with optimal dilutions of commercially prepared antibodies. Reagents used were CD3ε PE (145-C11), CD11b PE (M1/70), CD45R PE (RA3-6B2), CD117 eFluor780 (2B8), Sca-1 PerCP-Cy5.5 (D7; eBiosciences), CD4 PE (H129.19), CD8a PE (53–6.7), CD19 PE (1D3), Gr-1 PE (RB6-8C5), NK1.1 PE (PK136), and Ter119 PE (Ter119; BD Pharmingen). The lineage cocktail was composed of CD3ε, CD4, CD8a, CD11b, CD19, CD45R, Gr-1, NK1.1, and Ter119. Dead cells were excluded by staining with Sytox Blue (Invitrogen). Approximately 5,000 LSK cells were sorted by FACSAriaIII (Becton Dickinson) and injected intravenously into nonirradiated triple transgenic Rag2^−/−γ_c^−/−Kit^W/Wv recipient mice. Donor chimerism of blood cells was determined 1 mo after transplantation.

Singhal M., Liu X., Inverso D., Jiang K., Dai J., He H., Bartels S., Li W., Abdul Pari A.A., Gengenbacher N., Besemfelder E., Hui L., Augustin H.G, & Hu J. (2018). Endothelial cell fitness dictates the source of regenerating liver vasculature. The Journal of Experimental Medicine, 215(10), 2497-2508.

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