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Igd fitc

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IgD-FITC is a fluorescently labeled antibody that binds to the IgD isotype of immunoglobulins. It is designed for use in flow cytometry and other immunoassay applications.

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Lab products found in correlation

Cd27 pe cy7, by BD (4 mentions) Lsrfortessa, by BD (4 mentions) Facsdiva software, by BD (4 mentions) Cd24 pe, by BD (3 mentions) Cd45ra pe, by BD (3 mentions) Ccr7 pe, by BioLegend (3 mentions) Cd27 pe, by BD (3 mentions) Cd21 pe cy5, by BD (2 mentions) Ifn γ pe cy7, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Igm pe, by BD (2 mentions) Cd4 apc, by BD (2 mentions) Cd45 bv510, by BD (2 mentions) Histopaque 1077, by Merck Group (2 mentions) Cd45ra percp cy5, by Thermo Fisher Scientific (2 mentions) Cd19 bv450, by BD (2 mentions) Cd62l apc ef780, by Thermo Fisher Scientific (2 mentions) Cxcr5 af488, by BioLegend (2 mentions) Hla dr efluor450, by Thermo Fisher Scientific (2 mentions) Cxcr3 fitc, by BioLegend (2 mentions)

Spelling variants of this product

Anti-IgD-FITC (22 citations) IgD-FITC (IA6-2, (4 citations) FITC-conjugated anti-IgD (clone 11-26c.2a, (2 citations) Anti-IgD-FITC (clone IA6-2, (2 citations) α-IgD-FITC (2 citations) Anti-IgD-FITC (IA6-2, (2 citations) Human IgD-FITC (2 citations)

22 protocols using igd fitc

1

Multiparameter Flow Cytometry Analysis

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After 6 days culture, cultured cells were analyzed for expression of surface markers by multi-color flow cytometry. Cell aliquots were treated with anti-human Fc mAb for 20 minutes and stained for 30 minutes with selected combinations of fluorochrome-conjugated antibodies.
For the analysis of B cell subpopulations and activation markers, the monoclonal antibodies used were CD19-PE-CF594 (BD Biosciences, San Jose, CA), CD62L-PE-Cy5 (BD Biosciences), IgD-FITC (BD Biosciences), CD27-PE-Cy7 (BD Biosciences), CD24-PE (BD Biosciences), CD38-PerCP-Cy5.5 (Beckman Coulter, Brea CA), CD138-PE-Cy7 (eBioscience, San Diego, CA), CD23-PE (BD Biosciences), CD21-PE-Cy5 (BD Biosciences), IgG-PC7 (BD Biosciences), CD86-Alexa 700 (BD Biosciences) and CD95-Pacific Blue (BioLegend, San Diego, CA).
For plasma cell staining, cultured cells were first incubated with monoclonal antibodies to surface markers (CD38, CD138 and CD19), fixed and permeabilized with Cytofix/Cytoperm Fixation/Permeabilization Solution Kit (BD Biosciences) and then incubated with Blimp-1-PE (R&D systems) and Pax5-APC (eBioscience) for 30 minutes.
For the proliferation assay, 1–10×106 cells of interest were labeled with 1μM carboxyfluorescein succinimidyl ester (CFSE) (Invitrogen) before culture initiation. After 6 days of culture, cells were harvested, incubated with antibodies, and dilution of CFSE was assessed by flow cytometry.
Traitanon O., Mathew J.M., La Monica G., Xu L., Mas V, & Gallon L. (2015). Differential Effects of Tacrolimus versus Sirolimus on the Proliferation, Activation and Differentiation of Human B Cells. PLoS ONE, 10(6), e0129658.
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2

Flow Cytometry of Immune Cell Subsets

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The following fluorochrome conjugated antihuman monoclonal antibodies (MoAbs) were used for flow cytometry studies: ICOSBV421, ICOSAlexa488, CD40LBV605, CD69BV650, HLA-DRFITC, CD38APCCy7, and TNF-αAPCCy7 from BioLegend (San Diego, CA); CD3BUV395, CD4PerCPCy5.5, CD8Alexa-Fluor700, CCR7PECF594, IL-2BV711, CXCR5Alexa647, IFNγPE-Cy7, PD1BV650, CD45ROAPCH7, CD21PECy5, CD27PerCPCy5.5, IgDFITC, CD10PECy7, and CD20Alexa700 from BD Bioscience (San Jose, CA); IL-21PE, CD27PECy5, and IL-17Alexa488 from e-Biosciences (San Diego, CA); and CD45ROPE-TexasRed from Beckman Coulter (Fullerton, CA). Live/Dead Fixable Aqua Dead Cell Stain Kit and CellTrace Violet Cell Proliferation Kit were from ThermoFisher (Boston, MA). Recombinant human IL-21 (Cat #8879-IL-010) from R&D systems, purified antihuman IL-2 (Cat #3440-ON-500) and antihuman-IL-21 (Cat #MT216G/21.3m) from Mabtech, and antihumanTNF-α (Cat #502922) from BioLegend were used. Samples were acquired on a BD LSRFortessa (BD Biosciences, CA) flow cytometer and analyzed by FlowJo V10 (TreeStar, Inc).
Pallikkuth S., de Armas L.R., Rinaldi S., George V.K., Pan L., Arheart K.L., Pahwa R, & Pahwa S. (2019). Dysfunctional peripheral T follicular helper cells dominate in people with impaired influenza vaccine responses: Results from the FLORAH study. PLoS Biology, 17(5), e3000257.
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3

Analysis of B-cell Subsets by Flow Cytometry

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To analyze the B-cell subsets, PBMCs were stained with fluorochrome-labelled anti-human CD10-APC, CD19-APC-Cy7, CD27-PC7, CD38-PercCP-Cy5.5, IgD-FITC, and IgM-PE (BD Biosciences, San Jose, CA, USA). Antibody-stained cells were analyzed by flow cytometry using FACSCANTOII and Diva software (BD Biosciences). The gating strategy is outlined in Fig 1A. The absolute numbers of subsets were obtained by multiplying their percentage by the total lymphocyte number.
Aspord C., Bruder Costa J., Jacob M.C., Dufeu-Duchesne T., Bertucci I., Pouget N., Brevot-Lutton O., Zoulim F., Bourliere M., Plumas J, & Leroy V. (2016). Remodeling of B-Cell Subsets in Blood during Pegylated IFNα-2a Therapy in Patients with Chronic Hepatitis B Infection. PLoS ONE, 11(6), e0156200.
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4

Isolation and Characterization of PBMCs

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Peripheral blood mononuclear cells (PBMCs) were obtained from healthy individuals homozygous for the FcγRIIB-T232 or FcγRIIB-I232 site, under appropriate ethics approval from the NIHR Cambridge Bioresource. Inclusion criteria for individuals were people aged between 44 and 77 years, with no serious co-morbidities, no direct family history of autoimmune disease, no use of immunosuppressants or steroids and no hospitalisation within the last 12 months. Individuals were age and sex matched (18 females and 11 males matched between genotypes). Flow sorting was performed using CD19-BV785, CD38-BV711, CD3-NC650, CD14-605NC, CD24-PerCP-Cy5.5, IgD-FITC, CD27-PE-Cy7 (all from BD Bioscience) and Aqua (for live-dead cell detection, Invitrogen), where flow protocol is outlined in Supplementary Fig. 7.
Espéli M., Bashford-Rogers R., Sowerby J.M., Alouche N., Wong L., Denton A.E., Linterman M.A, & Smith K.G. (2019). FcγRIIb differentially regulates pre-immune and germinal center B cell tolerance in mouse and human. Nature Communications, 10, 1970.
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5

Comprehensive Flow Cytometry Immunophenotyping

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Cells were stained using Abs specific for mouse Ags: CD45R (B220) (various fluorochromes) (BD Pharmingen, BioLegend, San Diego, CA and Tonbo Biosciences); IgM (various fluorochromes, Jackson ImmuoResearch Laboratories, West Grove, PA); CD19 eFlour450, CD25 APC, MHC II APC (Tonbo Biosciences); CD43 PE, BP-1 PE, CD117 PE, CD24 (HSA) PE-Cy7, IgD FITC (BD Pharmingen); CD21 PerCP/Cy5.5 (BioLegend). IC staining was performed with IC fixation and permeabilization buffer (eBiosciences). Methanol fixation was performed for IC phospho (p)-S6R detection. Abs used for IC staining were p-ribosomal S6 protein (S6R) S235/236 PE (eBiosciences); p-AMPKα T172, c-Myc AF488 and p-Ulk1 S555 (Cell Signaling Technology, Danvers, MA). Donkey anti-rabbit Alexa-Fluor 647 (Life Technologies, Carlsbad CA) secondary Ab was used to detect unlabeled primary Abs. Data was collected using FACS Canto II or LSR II flow cytometers (BD Biosciences) and analyses were performed using FlowJo software (TreeStar, Ashland, OR).
Ramίrez J.A., Iwata T., Park H., Tsang M., Kang J., Cui K., Kwong W., James R.G., Baba M., Schmidt L.S, & Iritani B.M. (2019). Folliculin Interacting Protein-1 Maintains Metabolic Homeostasis During B Cell Development by Modulating AMPK, mTORC1, and TFE3. Journal of immunology (Baltimore, Md. : 1950), 203(11), 2899-2908.
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6

Comprehensive Immune Cell Profiling

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The following antibodies were used: CD4-AlexaFluor 700, CXCR5-Biotin, ICOS-PECy7, Bcl-6-AlexaFluor 647, CD27-APCH7, CD45RA-PE, CD19-APC or V450, IgD-FITC, IL-21-AlexaFluor 647, IFN-γ-PECy7, IL-10-PE, Streptavidin-PE-TexasRed (BD Biosciences), CD38-PerCPeFluor710, IL-6-FITC, (Biolegend). Neutralizing antibodies specific for human IL-6 and IL-21R and isotype controls from R&D Systems.
Chavele K.M., Merry E, & Ehrenstein M.R. (2015). Cutting Edge: Circulating Plasmablasts Induce the Differentiation of Human T Follicular Helper Cells via IL-6 Production. The Journal of Immunology Author Choice, 194(6), 2482-2485.
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7

Multiparametric Flow Cytometry Immunophenotyping

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Cell surface staining was processed using whole blood lysis method. Freshly drawn heparinized peripheral blood samples were labeled with monoclonal antibody (mAb) panel for T lymphocyte subsets [anti-CD3-AlexaFlour700, -CD4-APC, -CD8-APC/CY7, -CD45-BV510, -CD45RA-FITC, -CD45RO-PE, -CCR7-PE and -HLA-DR-BV711 (all from Biolegend, San Jose, CA, USA)], mAb panel for B lymphocyte subsets [anti-CD19-APC, -CD24-PE, -CD27-PE/CY7, -CD38-AlexaFlour700, -CD45-BV510, -CD138-BV421 and -IgD-FITC (all from BD-Biosciences, San Jose, CA, USA)], and with mAb panel for Treg [anti-CD3-AlexaFlour700, -CD4-APC, -CD25-PE, -CD45-BV510, -CD127-BV421 (all from BD-Biosciences, San Jose, CA, USA)]. Following incubation, erythrocytes were lysed using FACS Lysing Solution (BD Biosciences, San Jose, CA, USA), and at least 10.000 cells were collected in CD45+ lymphocyte gate. The data were acquired on a NovoCyte flow cytometer (Agilent Technologies, USA) and analyzed using the NovoExpress operating system software (Agilent Technologies, USA).
Gelmez M.Y., Oktelik F.B., Tahrali I., Yilmaz V., Kucuksezer U.C., Akdeniz N., Cetin E.A., Kose M., Cinar C., Oguz F.S., Besisik S., Koksalan K., Ozdemir O., Senkal N., Gul A., Tuzun E, & Deniz G. (2022). Immune modulation as a consequence of SARS-CoV-2 infection. Frontiers in Immunology, 13, 954391.
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8

Multiparametric Flow Cytometry Immunophenotyping

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Peripheral blood mononuclear cells were prepared for analysis by density centrifugation using Histopaque-1077 (Sigma-Aldrich). The following antibodies were used for flow cytometry immunophenotyping: CD3 – BV605 (Biolegend, San Diego, CA, USA), CD4 – APC-eFluor780 (eBioscience, San Diego, CA,USA), CD8 – BV650 (eBioscience, San Diego, CA,USA), CD25 – PE (eBioscience, San Diego, CA,USA), CD127 – APC (eBioscience, San Diego, CA,USA), CD45RA – PerCP-Cy5.5(eBioscience, San Diego, CA,USA, CD19 – BV450 (BD Bioscience, Franklin Lakes, NJ, USA), CD27 – PE-Cy7 (eBioscience, San Diego, CA,USA, CD62L – APC-eF780 (eBioscience, San Diego, CA,USA, CXCR3 – FITC (Biolegend, San Diego, CA, USA), CXCR5 – AF488 (Biolegend, San Diego, CA, USA), CCR7 – PE (Biolegend, San Diego, CA, USA), PD-1 – APC (eBioscience, San Diego, CA,USA), HLA-DR- eFluor450 (eBioscience, San Diego, CA, USA), IgD – FITC (BD Bioscience, Franklin Lakes, NJ, USA). Flow cytometry analysis was performed on a BD LSRFortessa (BD Bioscience) with FACS Diva software (BD Bioscience) for acquisition, then analysis was performed with FlowJo software (LLC).
Thaventhiran J.E., Allen H.L., Burren O.S., Rae W., Greene D., Staples E., Zhang Z., Farmery J.H., Simeoni I., Rivers E., Maimaris J., Penkett C.J., Stephens J., Deevi S.V., Sanchis-Juan A., Gleadall N.S., Thomas M.J., Sargur R.B., Gordins P., Baxendale H.E., Brown M., Tuijnenburg P., Worth A., Hanson S., Linger R., Buckland M.S., Rayner-Matthews P.J., Gilmour K.C., Samarghitean C., Seneviratne S.L., Sansom D.M., Lynch A.G., Megy K., Ellinghaus E., Ellinghaus D., Jorgensen S.F., Karlsen T.H., Stirrups K.E., Cutler A.J., Kumararatne D.S., Chandra A., Edgar J.D., Herwadkar A., Cooper N., Grigoriadou S., Huissoon A., Goddard S., Jolles S., Schuetz C., Boschann F., Lyons P.A., Hurles M.E., Savic S., Burns S.O., Kuijpers T.W., Turro E., Ouwehand W.H., Thrasher A.J, & Smith K.G. (2020). Whole genome sequencing of a sporadic primary immunodeficiency cohort. Nature, 583(7814), 90-95.
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9

Immunophenotyping of B Cell Subsets

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The immunophenotyping of B cells was performed in the PB samples (50 μl) using the following fluorochrome-labelled antibodies: CD19-BB700, CD27-APC, IgD-FITC, CD5-FITC (all BD Biosciences), and TLR10-PE (BioLegend). Fresh whole blood was stained with antibodies for 20 minutes at room temperature (RT) in the dark, lysed with 500 μl of lysis buffer (BD) for 15 minutes under the same conditions, washed twice with PBS, resuspended, and analysed by flow cytometry. Approximately 100,000 events were collected per sample. The data were collected with a FACSCalibur (BD Bioscience) and analysed using the FlowJo software version 10.0. Figure 1 shows the results for CD19+ B cells and the subset gating strategy used in these experiments.
Zhang Y., Cao R., Ying H., Du J., Chen S., Wang N, & Shen B. (2018). Increased Expression of TLR10 in B Cell Subsets Correlates with Disease Activity in Rheumatoid Arthritis. Mediators of Inflammation, 2018, 9372436.
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10

Lymphocyte Subset Analysis by Flow Cytometry

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Flow cytometry was used to measure the percentages of lymphocyte populations in whole blood using the monoclonal antibody conjugates (6 Color TBNK Reagent, CD3-PerCP-Cy5.5, CD4-APC, CD8-V450, CCR7-FITC, CD45RA-PE, CD19-PerCP-Cy5.5, CD27-PE, IgD-FITC, CD24-APC and CD38V450, all from BD Biosciences (Franklin Lakes NJ). The percentage of each patient’s lymphocyte subset was compared with normal control ranges for age that have either been established independently in the Boston Children’s flow cytometry laboratory. Lymphocyte proliferation assays were performed in three independent experiments.
Frugoni F., Dobbs K., Felgentreff K., Aldhekri H., Al Saud B.K., Arnaout R., Ali A.A., Abhyankar A., Alroqi F., Giliani S., Ojeda M.M., Tsitsikov E., Pai S.Y., Casanova J.L., Notarangelo L.D, & Manis J.P. (2015). A Novel Mutation in the POLE2 Gene Causing Combined Immunodeficiency. The Journal of allergy and clinical immunology, 137(2), 635-638.e1.
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