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Anti cd127

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Anti-CD127 is a monoclonal antibody that binds to the CD127 (IL-7 receptor alpha) cell surface antigen. It is commonly used in flow cytometry applications to identify and analyze CD127-expressing cells.

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Lab products found in correlation

Lsrfortessa, by BD (6 mentions) Anti cd45, by BioLegend (4 mentions) Anti cd11b, by BioLegend (4 mentions) Anti sca 1, by Thermo Fisher Scientific (3 mentions) Anti cd28, by Thermo Fisher Scientific (3 mentions) Anti cd57, by Thermo Fisher Scientific (3 mentions) Facsaria 3, by BD (2 mentions) Tissuelyser 2, by Qiagen (2 mentions) Collagenase type 4, by Worthington (2 mentions) Annexin 5 af647, by BioLegend (2 mentions) Trustain fcx, by BioLegend (2 mentions) Anti siglec f, by Thermo Fisher Scientific (2 mentions) Anti b220, by Thermo Fisher Scientific (2 mentions) Anti cd34, by Thermo Fisher Scientific (2 mentions) Anti cd16 32, by Thermo Fisher Scientific (2 mentions) Zombie nir fixable viability kit, by BioLegend (2 mentions) Anti gr 1, by BD (2 mentions) Propidium iodide, by BioLegend (2 mentions) Anti cd3, by BioLegend (2 mentions) Anti ly6g, by BD (2 mentions)

Spelling variants of this product

Anti-CD127-PE-Cy7 (9 citations) Anti-CD127 (A7R34) (5 citations) Anti-CD127 FITC (4 citations) Anti-CD127 PE-Cy5 (clone eBioRDR5, (2 citations) Anti-CD127-PE (2 citations) PE-Cy5-conjugated anti-CD127 (clone A7R34; (2 citations) EFour450-conjugated anti-CD127 (2 citations) Anti-CD127 biotin (clone A7R34, (2 citations) PE-Cy7-conjugated anti-CD127 (2 citations) Anti-CD127 (A7R34) antibody (2 citations)

14 protocols using anti cd127

1

Isolation and Flow Cytometry of Neutrophils

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Flow cytometry analysis was performed on a LSRII or LSRFortessa (BD Biosciences). To isolate neutrophils from the feet, tissue was diced and incubated at 37°C for 45 min with shaking at 200 rpm (TissueLyserII, Qiagen), in DMEM containing 1 mg/mL Collagenase Type IV (Worthington Biochemical). Cells were then strained through a 70 μM strainer and washed twice in phosphate buffered saline (PBS). Neutrophils from bone marrow and feet were sorted on a FACSAria III (BD Biosciences). FcγR were blocked using TruStain FcX (BioLegend). Data were analyzed using FlowJo software (BD Biosciences). The following reagents were used: Zombie-NIR fixable viability kit (BioLegend), anti-B220 (eBioscience;RA36B2), anti-CD3 (WEHI;KT3.1.1), anti-CD4 (WEHI;GK1.5), anti-CD8 (WEHI;53.6.7), anti-CD11b (BioLegend;M1/70), anti-CD16/32 (WEHI;24G2), anti-CD34 (eBioscience;RAM34), anti-CD45.1 (BioLegend;A20), anti-CD45.2 (BioLegend;104), anti-CD48 (BD Bioscience;C2), anti-CD127 (eBioscience;A7R34), anti-CD135 (BioLegend;A2F10), anti-CD150 (BioLegend;TC15–12F12.2), anti-cKit (WEHI;ACK4), anti-Gr1 (WEHI;RB6–8C5), anti-Ly6G (BD;1A8), anti-Sca-1 (WEHI;Ly6A/E), anti-Siglec-F (Fisher;1RMN44N), AnnexinV-AF647 (BioLegend), and propidium iodide (Sigma).
Speir M., Nowell C.J., Chen A.A., O’Donnell J.A., Shamie I.S., Lakin P.R., D’Cruz A.A., Braun R.O., Babon J.J., Lewis R.S., Bliss-Moreau M., Shlomovitz I., Wang S., Cengia L.H., Stoica A.I., Hakem R., Kelliher M.A., O’Reilly L.A., Patsiouras H., Lawlor K.E., Weller E., Lewis N.E., Roberts A.W., Gerlic M, & Croker B.A. (2019). Ptpn6 inhibits caspase-8- and Ripk3/Mlkl-dependent inflammation. Nature immunology, 21(1), 54-64.
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2

Characterizing IL-1β-producing Lung Cells

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Lungs from sham- and RV-treated immature C57BL/6J or IL-1β−/− mice were harvested one or seven days post-infection, perfused with PBS containing EDTA, minced, and digested in collagenase IV. Cells were filtered and washed with RBC lysis buffer, and dead cells were stained with PacBlue (Thermo Fisher Scientific). To identify the cellular source of IL-1β, lung cells were harvested one day post-infection and stained with fluorescent-tagged anti-CD45, anti-F4/80, and anti-CD11b (all from BioLegend). Cells were subsequently treated with permeabilization buffer (eBioscience) and stained with anti-IL-1β (eBioscience). To identify ILC2s, cells were then stained with fluorescent-tagged antibodies for lineage markers (CD3ε, TCRb, B220/CD45R,Ter-119, Gr-1/Ly-6G/Ly-6C, CD11b, CD11c, F4/80, and FcεRIa; all from BioLegend), anti-CD25 (BioLegend), and anti-CD127 (eBioscience), as described9 (link). Cells were fixed, subjected to flow cytometry, and analyzed on an LSR Fortessa (BD Biosciences, San Jose, CA). Data were collected using FACSDiva software (BD Biosciences) and analyzed using FlowJo software (TreeStar, Ashland, OR).
Han M., Ishikawa T., Bermick J.R., Rajput C., Lei J., Goldsmith A.M., Jarman C.R., Lee J., Bentley J.K, & Hershenson M.B. (2020). IL-1β prevents ILC2 expansion, type 2 cytokine secretion and mucus metaplasia in response to early-life rhinovirus infection in mice. Allergy, 75(8), 2005-2019.
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3

Comprehensive Immunostaining Panel for Immune Cell Analysis

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Anti-Nono (Cat# 11058-1-AP and 66361-1-Ig) was from Proteintech. Anti-Elk3 (Cat# NBP1-83960) was from Novus Biologicals. Anti-Tmem241 (Cat# 203644-T32) was from Sinobiological. Anti-Lineage cocktail (Cat# 88-7772-72), Anti-CD127 (A7R34), anti-Sca-1 (D7), anti-Flt3 (A2F10), anti-α4β7 (DATK32), anti-Id2 (ILCID2), anti-PLZF (Mags.21F7), anti-Eomes (Dan11mag), anti-NKp46 (29A1.4), anti-NK1.1 (PK136), anti-CD45 (30-F11), anti-CD25 (PC61.5), anti-Gata3 (TWAJ), anti-RORγt (AFKJS-9), anti-Bcl11b (1F8H9), anti-CD3 (17A2), anti-CD19 (1D3), anti-KLRG1 (2F1), anti-CD90 (HIS51), anti-IL-22 (IL22JOP), anti-Ki67 (SolA15), anti-CD45.2 (104), anti-BrdU (BU20A), anti-PD-1 (J43) and anti-CD45.1 (A20) were purchased from eBiosciences (San Diego, USA). Anti-c-Kit (2B8), anti-CD150 (TC15-12F12.2), anti-CD48 (HM48-1), and anti-CD49a (HMα1) were purchased from Biolegend (California, USA). Anti-β-actin (Cat# RM2001) was purchased from Beijing Ray Antibody Biotech. Paraformaldehyde (PFA) and 4’,6-diamidino-2-phenylindole (DAPI) were from Sigma. IL-22 ELISA kit was purchased from Neobioscience.
Liu N., He J., Fan D., Gu Y., Wang J., Li H., Zhu X., Du Y., Tian Y., Liu B, & Fan Z. (2022). Circular RNA circTmem241 drives group III innate lymphoid cell differentiation via initiation of Elk3 transcription. Nature Communications, 13, 4711.
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4

Isolation and Flow Cytometry of Neutrophils

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Flow cytometry analysis was performed on a LSRII or LSRFortessa (BD Biosciences). To isolate neutrophils from the feet, tissue was diced and incubated at 37°C for 45 min with shaking at 200 rpm (TissueLyserII, Qiagen), in DMEM containing 1 mg/mL Collagenase Type IV (Worthington Biochemical). Cells were then strained through a 70 μM strainer and washed twice in phosphate buffered saline (PBS). Neutrophils from bone marrow and feet were sorted on a FACSAria III (BD Biosciences). FcγR were blocked using TruStain FcX (BioLegend). Data were analyzed using FlowJo software (BD Biosciences). The following reagents were used: Zombie-NIR fixable viability kit (BioLegend), anti-B220 (eBioscience;RA36B2), anti-CD3 (WEHI;KT3.1.1), anti-CD4 (WEHI;GK1.5), anti-CD8 (WEHI;53.6.7), anti-CD11b (BioLegend;M1/70), anti-CD16/32 (WEHI;24G2), anti-CD34 (eBioscience;RAM34), anti-CD45.1 (BioLegend;A20), anti-CD45.2 (BioLegend;104), anti-CD48 (BD Bioscience;C2), anti-CD127 (eBioscience;A7R34), anti-CD135 (BioLegend;A2F10), anti-CD150 (BioLegend;TC15–12F12.2), anti-cKit (WEHI;ACK4), anti-Gr1 (WEHI;RB6–8C5), anti-Ly6G (BD;1A8), anti-Sca-1 (WEHI;Ly6A/E), anti-Siglec-F (Fisher;1RMN44N), AnnexinV-AF647 (BioLegend), and propidium iodide (Sigma).
Speir M., Nowell C.J., Chen A.A., O’Donnell J.A., Shamie I.S., Lakin P.R., D’Cruz A.A., Braun R.O., Babon J.J., Lewis R.S., Bliss-Moreau M., Shlomovitz I., Wang S., Cengia L.H., Stoica A.I., Hakem R., Kelliher M.A., O’Reilly L.A., Patsiouras H., Lawlor K.E., Weller E., Lewis N.E., Roberts A.W., Gerlic M, & Croker B.A. (2019). Ptpn6 inhibits caspase-8- and Ripk3/Mlkl-dependent inflammation. Nature immunology, 21(1), 54-64.
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5

Characterization of Senescent and Exhausted T Cells

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PBMCs were isolated using Ficoll-Paque (GE Healthcare, USA), and the cellular phenotypic of senescent and exhausted T cells was analyzed by flow cytometry. For surface markers analysis, cells were stained in PBS containing 2% fetal bovine serum (FBS, Thermo fisher, USA) with antibodies as indicated. Then, flow cytometric analysis was carried out in BD LSRFortessa X20. The gating strategy to identify T cell subsets was applied as described previously (27 (link)). Antibodies used in this study were purchased from BD Biosciences and eBioscience, including anti-CD4 (GK1.5, 1:100), anti-CD8 (53-6.7, 1:100), anti-CD25 (PC61, 1:100), anti-CD45RA (HI100, 1:100), anti-CXCR3(G025H7, 1:100), anti-CCR4 (L291H4, 1:100), anti-CCR6 (G034E3, 1:100), anti-CCR7 (G043H7, 1:100), anti-CD127 (A019D5, 1:100), anti-CXCR5 (RF8B2, 1:100), anti-CD28 (CD28.2, 1:100), anti-CD57 (NK-1, 1:100), anti-KLRG1 (2F1, 1:100), anti-PD-1 (EH12.2H7, 1:100), anti-TIM3 (F38-2E2, 1:100), and fixable viability dye eFluor 780 (eBioscience, Cat#65-0865-14, 1:1,000).
Tang X., Deng B., Zang A., He X., Zhou Y., Wang D., Li D., Dai X., Chen J., Zhang X., Liu Y., Xu Y., Chen J., Zheng W., Zhang L., Gao C., Yang H., Li B, & Wang X. (2022). Characterization of age-related immune features after autologous NK cell infusion: Protocol for an open-label and randomized controlled trial. Frontiers in Immunology, 13, 940577.
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6

Comprehensive Immune Cell Profiling by Flow Cytometry

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Cell surface staining were performed with the following antibodies: anti-CD3ε, and anti-CD4, anti-CD11b, anti-CD11c, anti-CD19, anti-B220, and anti-Ly-6G (TONBO Bioscience, San Diego, CA, USA); anti-IL-33R (ST2), anti-TCRγδ, anti-CD90.2, and anti-CD45 (BioLegend, San Diego, CA, USA); anti-TCRβ, anti-CD25, anti-CD44, anti-CD62L, anti-CD69, anti-CD103, anti-NK1.1, and anti-TER119 (BD Bioscience); and anti-CD127 (eBioscience, San Diego, CA, USA). The same procedures were conducted with the following antibodies for the intracellular staining: anti-IL-4 and anti-IL-5 (BD Bioscience); and anti-IL-13 (eBioscience). Fixable Viability Dye (eBioscience) was used to exclude dead cells. All samples were analyzed using the FACS Verse (BD Bioscience) and Flow Jo software programs (Tree Star Inc., San Carlos, CA, USA).
Mato N., Hirahara K., Ichikawa T., Kumagai J., Nakayama M., Yamasawa H., Bando M., Hagiwara K., Sugiyama Y, & Nakayama T. (2017). Memory-type ST2+CD4+ T cells participate in the steroid-resistant pathology of eosinophilic pneumonia. Scientific Reports, 7, 6805.
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7

Identification of Lung ILC2s in Mice

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Lungs from sham- and RV-treated immature wild-type BALB/c or TSLPR KO mice were perfused with PBS containing EDTA, minced and digested in collagenase IV. Cells were filtered and washed with RBC lysis buffer, and dead cells were stained with Pac-Orange Live/Dead fixable dead staining dye (Invitrogen). To identify ILC2s, cells were then stained with fluorescent-tagged antibodies for lineage markers (CD3ε, TCRβ, B220/CD45R, Ter-119, Gr-1/Ly-6G/Ly-6C, CD11b, CD11c, F4/80 and FcεRIα, all from Biolegend), anti-CD25 (Biolegend), and anti-CD127 (eBioscience), as described (21 (link)). Cells were fixed, subjected to flow cytometry and analyzed on a LSR Fortessa (BD Biosciences, San Jose, CA). Data were collected using FACSDiva software (BD Biosciences) and analyzed using FlowJo software (Tree Star, Ashland, OR).
Han M., Rajput C., Hong J.Y., Lei J., Hinde J.L., Wu Q., Bentley J.K, & Hershenson M.B. (2017). The innate cytokines IL-25, IL-33 and TSLP cooperate in the induction of ILC2 expansion and mucous metaplasia in rhinovirus-infected immature mice. Journal of immunology (Baltimore, Md. : 1950), 199(4), 1308-1318.
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8

Comprehensive Multiparametric Flow Cytometry Analysis

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PBMCs were incubated with fluorochrome-conjugated mAbs and analyzed on either an FC 500 (Beckman Coulter) or an LSRFortessa (Becton Dickinson) flow cytometer. FITC-, PE-, PE-Texas Red-, ECD-, APC-, PE-Cy5–, PE-Cy7–, PerCP-Cy5.5–, Pacific Blue-, and AF700-conjugated mouse anti-human mAbs included anti-CD3, anti-CD4, anti-CD8, anti-CD11c, anti-CD14, anti-CD16 (clone 3G8), anti-CD19, anti-CD25, ant-CD28, anti-CD45RA, anti-CD45RO, anti-CD80, anti-CD86, anti-CD123, anti-CTLA-4, anti–HLA-DR, anti-IL2, anti-Ki-67 (BD Pharmingen), anti-CD56, anti-CD83 (Beckman Coulter), anti-CD127, anti- IFNγ, anti-LAG-3, anti-PD-1, anti- TNFα (eBioscience), anti-CCR7, anti-TIM-3 (R&D Systems), and anti-CD57 (BioLegend). Nonreactive isotype-matched antibodies (Becton Dickinson, eBioscience, R&D Systems) were used as controls. LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Life Technologies) facilitated exclusion of dead cells. Gates were set for collection and analysis of at least 20,000 live events. Data were analyzed with FlowJo 9.5 software (TreeStar).
Chung D.J., Pronschinske K.B., Shyer J.A., Sharma S., Leung S., Curran S.A., Lesokhin A.M., Devlin S.M., Giralt S.A, & Young J.W. (2015). T cell exhaustion in multiple myeloma relapse after autotransplant: Optimal timing of immunotherapy. Cancer immunology research, 4(1), 61-71.
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9

Enrichment and Characterization of Human Immune Cells

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The human prostate cancer cell line, PC3 (kindly provided by Dr. Reiter, UCLA), was cultured with RPMI 1640 medium (Mediatech, Manassas, VA) supplemented with 10% FBS (Omega Scientific, Tarzana, CA), and penicillin–streptomycin–fungizone (Life Technologies, Carlsbad, CA). Human cells were cultured in RPMI 1640 supplemented with 10% human AB serum (Omega Scientific), penicillin–streptomycin–fungizone, and 10 mM HEPES (Life Technologies). Monoclonal antibody used for human T-and B-cell enrichment included anti-CD14, anti-CD16, and anti-CD25, and for DC enrichment included anti-CD3, anti-CD16, and anti-CD19, all from BD Biosciences (San Jose, CA). Anti-mouse Ig-conjugated immunomagnetic beads were from Life Technologies (M450 Dynabeads; Carlsbad, CA). Fluorochrome-conjugated mAbs against human CD3, CD4, CD8, CD56, CD69, and CD45RA were obtained from BD Biosciences; anti-CD127 from eBiosciences (San Diego, CA); and anti-CD25 from BioLegend (San Diego, CA,).
Roth M.D, & Harui A. (2015). Human tumor infiltrating lymphocytes cooperatively regulate prostate tumor growth in a humanized mouse model. Journal for Immunotherapy of Cancer, 3, 12.
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10

Multiparametric Flow Cytometry of Immune Cells

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Fluorescent monoclonal antibodies (mAbs) were purchased as follows: anti-CD8, anti-CD56, anti-CD94, anti-NKG2D, anti-CTLA-4 and anti-TNF from BD Bioscience (San Jose, CA); anti-CD127, anti-CD3, anti-CD28 and anti-Eomesodermin (Eomes) from eBioscience (San Diego, CA); anti-PD-1, anti-Tbet and anti-IFNγ from BioLegend (San Diego, CA); anti-NKG2A from R&D Systems (Minneapolis, MN); anti-Vδ1 TCR (clone REA173) from Miltenyi Biotec (San Diego, CA); anti-Vδ2 TCR (clone B6), anti-Vγ9 TCR (clone B3) and pan-γδ TCR (clone B1) from BioLegend (San Diego, CA). Dead cells were excluded using Aqua dead cell stain kit (Life Technologies). The phycoerythrin- or allophycocyanin-labeled CD1d tetramers were kindly provided by the NIH Tetramer Facility at Emory University (Atlanta, GA).
Chang K.M., Traum D., Park J.J., Ho S., Ojiro K., Wong D.K., Wahed A.S., Terrault N.A., Khalili M., Sterling R.K., Janssen H.L., Shuhart M.C., Lau D.T., Roberts L.R., Johnson G.S., Kaplan D.E., Betts M.R., Lee W.M, & Lok A.S. (2019). Distinct phenotype and function of circulating Vδ1+ and Vδ2+ γδT-cells in acute and chronic hepatitis B. PLoS Pathogens, 15(4), e1007715.
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