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Anti cd3 fitc

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Anti-CD3-FITC is a fluorescently labeled antibody that binds to the CD3 surface marker on T cells. It can be used for the identification and enumeration of T cells in flow cytometry applications.

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Lab products found in correlation

Anti ifn γ pe, by Thermo Fisher Scientific (6 mentions) Anti foxp3 pe, by Thermo Fisher Scientific (6 mentions) Anti cd4 pe, by Thermo Fisher Scientific (4 mentions) Anti cd4 pe cy7, by Thermo Fisher Scientific (4 mentions) Anti cd8 apc, by Thermo Fisher Scientific (4 mentions) Anti cd19 pe, by Thermo Fisher Scientific (3 mentions) Facscalibur, by BD (3 mentions) Ionomycin, by Merck Group (3 mentions) Brefeldin a, by Merck Group (3 mentions) Anti cd4 percp, by Thermo Fisher Scientific (3 mentions) Anti cd4 fitc, by Thermo Fisher Scientific (3 mentions) Anti cd4 apc, by Thermo Fisher Scientific (3 mentions) Facscalibur flow cytometer, by BD (3 mentions) Anti cd25 pe, by Thermo Fisher Scientific (2 mentions) Anti cd45 v500, by BD (2 mentions) Anti cd38 pe cy7, by BD (2 mentions) Anti cd45ra fitc, by BD (2 mentions) Cd34 microbead kit, by Miltenyi Biotec (2 mentions) Anti cd34 percp efluor 710, by Thermo Fisher Scientific (2 mentions) Anti cd90 apc, by BD (2 mentions)

Spelling variants of this product

Anti-CD3 (512 citations) CD3-FITC (66 citations) Anti-mouse CD3-FITC (21 citations) FITC-conjugated anti-CD3 (15 citations) Anti-human CD3-FITC (10 citations) FITC conjugated anti-mouse CD3 (3 citations) Anti-CD3-FITC (UCHT1, (2 citations) FITC-labeled anti-CD3 (2 citations) Monoclonal anti-mouse CD3 (Pe-Cy5-labeled) and CD4 (FITC-labeled) antibodies (2 citations) Anti-chicken CD3-FITC (1 citations)

50 protocols using anti cd3 fitc

1

Comprehensive PBMC Immunophenotyping Protocol

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Cell counts were determined using a hematology analyzer (Sysmex XN‐330, Sysmex Co.). PBMCs were isolated using density‐gradient centrifugation (Ficoll‐Paque; GE Healthcare Bio‐Sciences AB) as previously described (Theall et al., 2020 ). Isolated PBMCs (1 × 106) were labeled with 100 μl of prediluted fluorochrome‐labeled antibodies (Abs). Combinations of Abs were used to characterize the T‐cell level of differentiation and nutrient‐sensing mechanism, namely anti‐CD3‐FITC (clone OKT3), anti‐CD69‐PE (clone FN50), anti‐CD25‐PE (clone BC96), anti‐CD71‐PE (clone OKT9), anti‐CD38‐PE (clone HB7), anti‐CD4‐PE (clone OKT4), anti‐CD57‐PE (clone TB01), anti‐CD4‐PErCP‐cy5.5 (clone RPA‐T4), anti‐CD8‐PerCP‐cy5.5 (clone RPA‐T8), anti‐CD36‐PerCP‐cy7 (clone eBioNL07), anti‐CD3‐APC (clone OKT3), and anti‐CD8‐APC (clone OKT8, Thermo Fisher Scientific); anti‐killer cell lectin‐like receptor G1 (KLRG1)‐FITC (clone REA261, Miltenyi Biotech); anti‐GLUT‐1‐PE (clone 202915, R&D Systems); anti‐GLUT‐4‐PE (Biorbyt). All mAbs were previously titrated to determine optimal dilutions for flow cytometry. Cells were incubated for 45‐min in the dark at room temperature.
Theall B., Stampley J., Cho E., Granger J., Johannsen N.M., Irving B.A, & Spielmann G. (2021). Impact of acute exercise on peripheral blood mononuclear cells nutrient sensing and mitochondrial oxidative capacity in healthy young adults. Physiological Reports, 9(23), e15147.
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2

Single-Cell Immunophenotyping of Immune Cells

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Single-cell suspensions from brains or PBMCs were stained with an antibody cocktail. The following antibodies were used: anti-CD3–FITC (11-0031, Thermo Fisher Scientific; 1:400), anti-CD8–eFluor 450 (48-0081, Thermo Fisher Scientific; 1:400), anti-CD122–PECy7 (25-1222, Thermo Fisher Scientific; 1:400), anti-CXCR3–BV510 (745033, BD Biosciences; 1:200), and rabbit anti–mouse ETGF (ab9585, Abcam; 1:400) subsequently bound with anti-rabbit–Alexa Fluor 488. Fluorochrome compensation was performed with single-stained OneComp eBeads (01-1111-41, Thermo Fisher Scientific). ImageStream analysis was performed using an Amnis Imaging Flow Cytometer (MilliporeSigma). Data analysis was performed using IDEAS software (MilliporeSigma).
Cai W., Shi L., Zhao J., Xu F., Dufort C., Ye Q., Yang T., Dai X., Lyu J., Jin C., Pu H., Yu F., Hassan S., Sun Z., Zhang W., Hitchens T.K., Shi Y., Thomson A.W., Leak R.K., Hu X, & Chen J. (2022). Neuroprotection against ischemic stroke requires a specific class of early responder T cells in mice. The Journal of Clinical Investigation, 132(15), e157678.
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3

Multiparameter Flow Cytometry for Myeloma

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Samples were washed in FACS Buffer (PBS with 2% FBS), incubated with Brilliant Buffer (BD) and FcR block (Miltenyi Biotec) for 5 minutes, and stained with either a Myeloma panel: multi-epitope anti-CD38-FITC (Cytognos) chosen to avoid masking from clinically administered antibodies and SAR442257 treatment ex vivo (Supplementary Fig. S1), anti-CD138-BV421 MI15, anti-CD45-BV510 HI30, anti-BCMA-PE 19F2 (BioLegend), anti-CD46-APC E4.3, anti-CD56-PerCP-Cy5.5 B159, anti-CD19-BV605 SJ25C1, anti-CD28-APC-R700 CD28.2, and LIVE/DEAD Fixable Near-IR Stain (Thermo Fisher Scientific) or non-multiple myeloma (non-MM) panel: anti-CD3-FITC, anti-CD4-BV421, anti-CD107a-PE H4A3, anti-CD107b-PE H4B4, anti-CD28- APC CD28.2, anti-CD38-PerCP-Cy5.5, anti-CD16-BV510, anti-CD8-BV605, anti-CD56-APC-R700 NCAM16.2, and LIVE/DEAD Fixable Near-IR Stain (Thermo Fisher Scientific) for 10 minutes. Multiple myeloma cells are classified as live, singlet, CD38+CD138+ and non-MM cells are gated as live, singlet, CD38−CD138− (Supplementary Fig. S1). All antibodies were from BD except when noted otherwise.
Keller A.L., Reiman L.T., Perez de Acha O., Parzych S.E., Forsberg P.A., Kim P.S., Bisht K., Wang H., van de Velde H, & Sherbenou D.W. (2024). Ex Vivo Efficacy of SAR442257 Anti-CD38 Trispecific T-cell Engager in Multiple Myeloma Relapsed After Daratumumab and BCMA-targeted Therapies. Cancer Research Communications, 4(3), 757-764.
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4

Flow Cytometry Immunophenotyping of Antibody-Secreting Cells

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Peripheral blood mononuclear cells were incubated with the following antibodies: anti-CD3 FITC, anti-CD19 PE-Cy7, anti-CD27 PE (Invitrogen) as well as anti-CD38 AF 647 and anti-CD20 PacBlue (Biolegend). Antibody secreting cells were gated as CD19+ CD3−, CD20low and then sub-gated as CD27highCD38high (LSRII-Blue), and data was analyzed using FlowJo software.
Cowan M., Chon W.J., Desai A., Andrews S., Bai Y., Veguilla V., Katz J.M., Josephson M.A., Wilson P.C., Sciammas R, & Chong A.S. (2014). Impact of Immunosuppression on Recall Immune Responses to Influenza Vaccination in Stable Renal Transplant Recipients. Transplantation, 97(8), 846-853.
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5

AML Immune Profiling Protocol

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Mononuclear cells (1×106/100 μl) from peripheral blood or bone marrow of AML patients were stained with the following antibodies (all from BD unless stated otherwise, dilution used and catalogue number in parentheses): anti-CD45RA-FITC (1:25, 555488), anti-CD90-APC (1:50, 561971), anti-CD135-Biotin (1:10, 624008), anti-CD38-PE-Cy7 (1:200, 335790), anti-CD10-Alexa-700 (1:10, 624040), anti-CD7-Pacific Blue (1:50, 642916), anti-CD45-V500 (1:200, 560777), anti-CD34-APC-Cy7 (1:100, custom made by BD, CD34 clone 581), anti-CD34-PerCP-Efluor 710 (1:100, e-Bioscience 46-0344-42), anti-CD33-PE-Cy5 (1:100, Beckman Coulter PNIM2647U), anti-CD19-PE (1:200, 349204), anti-CD3-FITC (1:100, 349201), anti-CD56-Alexafluor 647 (1:100, 557711), and Streptavidin-QD605 (1:200, Invitrogen Q10101MP). Samples from Patients #1, 10, 11 (remission sample only), 32, 35, and 55 were enriched for CD34+ cells using a Miltenyi CD34 MicroBead kit according to the manufacturer’s protocol prior to antibody staining. Cells were sorted on a FACS AriaIII to a post-sort purity of >95%.
Shlush L.I., Zandi S., Mitchell A., Chen W.C., Brandwein J.M., Gupta V., Kennedy J.A., Schimmer A.D., Schuh A.C., Yee K.W., McLeod J.L., Doedens M., Medeiros J.J., Marke R., Kim H.J., Lee K., McPherson J.D., Hudson T.J., Brown A.M., Trinh Q.M., Stein L.D., Minden M.D., Wang J.C, & Dick J.E. (2014). Identification of pre-leukemic hematopoietic stem cells in acute leukemia. Nature, 506(7488), 328-333.
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6

AML Immune Profiling Protocol

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Mononuclear cells (1×106/100 μl) from peripheral blood or bone marrow of AML patients were stained with the following antibodies (all from BD unless stated otherwise, dilution used and catalogue number in parentheses): anti-CD45RA-FITC (1:25, 555488), anti-CD90-APC (1:50, 561971), anti-CD135-Biotin (1:10, 624008), anti-CD38-PE-Cy7 (1:200, 335790), anti-CD10-Alexa-700 (1:10, 624040), anti-CD7-Pacific Blue (1:50, 642916), anti-CD45-V500 (1:200, 560777), anti-CD34-APC-Cy7 (1:100, custom made by BD, CD34 clone 581), anti-CD34-PerCP-Efluor 710 (1:100, e-Bioscience 46-0344-42), anti-CD33-PE-Cy5 (1:100, Beckman Coulter PNIM2647U), anti-CD19-PE (1:200, 349204), anti-CD3-FITC (1:100, 349201), anti-CD56-Alexafluor 647 (1:100, 557711), and Streptavidin-QD605 (1:200, Invitrogen Q10101MP). Samples from Patients #1, 10, 11 (remission sample only), 32, 35, and 55 were enriched for CD34+ cells using a Miltenyi CD34 MicroBead kit according to the manufacturer’s protocol prior to antibody staining. Cells were sorted on a FACS AriaIII to a post-sort purity of >95%.
Shlush L.I., Zandi S., Mitchell A., Chen W.C., Brandwein J.M., Gupta V., Kennedy J.A., Schimmer A.D., Schuh A.C., Yee K.W., McLeod J.L., Doedens M., Medeiros J.J., Marke R., Kim H.J., Lee K., McPherson J.D., Hudson T.J., Brown A.M., Trinh Q.M., Stein L.D., Minden M.D., Wang J.C, & Dick J.E. (2014). Identification of pre-leukemic hematopoietic stem cells in acute leukemia. Nature, 506(7488), 328-333.
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7

CD8+NKG2D+ Cells Analysis in Tumor

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For CD8+NKG2D+ cells analysis, cells were harvested and stained with anti-CD3-FITC (E10472-1633, eBioscience, USA), anti-CD8-PE (561946, BD, USA), anti-NKG2D-APC (130-099-216, Miltenyi Biotec, Germany), anti-CD27-PE (560985, BD, USA), and anti-CD57-APC (560845, BD, USA) antibodies. To detect the infiltration of CD8 or NK cells in the tumor mass, tumors were removed and crushed into single cell suspension. Then cells were stained with mouse anti-CD8-PErCp (46-0083-80, eBioscience, USA) or anti-NK1.1-FITC (553164, BD, USA) antibodies. To confirm the depletion of CD8 or NK cells, cells were isolated from peripheral blood and stained with anti-CD8-PErCp (46-0083-80) or anti-NK1.1-PerCp antibodies (45-5941-82) (all from eBioscience, USA). The staining was performed on ice for 30 min. Samples were acquired on the FACS Calibur (BD, USA) and data was analyzed using FlowJo software (Version 8.5.3, Tree Star Inc., USA).
Xu C., Lu X., Liu W., Chen A., Meng G., Zhang H., Li B., Zhang Y., Wu J, & Wei J. (2018). CD8+ T cells mediate the antitumor activity of frankincense and myrrh in hepatocellular carcinoma. Journal of Translational Medicine, 16, 132.
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8

Multiparametric Analysis of CD8+ T Cell Activation

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CD8+ T cells isolated as above were washed with FACS buffer (PBS containing 2% FBS and 2 mM EDTA) and labeled for 30 min in dark at 4 °C with the following monoclonal antibodies (TONBO biosciences) anti-CD3-FITC, anti-CD8a-PE, anti-TLR4-PE Cy7 (eBioscience, CA, USA), anti-CD28-PerCpCy5.5, anti-CD45RO-FITC(TONBO biosciences). After surface staining with anti-TLR4-PECy7 antibody, CD8+ T cells were then fixed for 30 min with 2% PFA, permeabilized with Perm buffer (TONBO biosciences) and labeled with anti-Granzyme B-PE (eBioscience), anti-Perforin-BV 421, anti-TNFα-PE, anti-IFNγ-APC/Alexa Fluor 700 antibodies (BD Biosciences). Expression levels were measured using BD LSRFortessa (BD Biosciences), and data were analyzed using FlowJo software (version 10; Tree Star). CD3+CD8+ T cells, directly gated in whole blood, (Supplementary Fig. 1b) were also analyzed for TLR4 surface protein expression.
Tripathy A., Khanna S., Padhan P., Smita S., Raghav S, & Gupta B. (2017). Direct recognition of LPS drive TLR4 expressing CD8+ T cell activation in patients with rheumatoid arthritis. Scientific Reports, 7, 933.
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9

Immune Cell Profiling in Atherosclerosis

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Splenocytes and blood were isolated from apoE-/-, catK-/-//apoE-/-, wt, and catK-/- mice. Erythrocytes in peripheral blood and spleen were removed by hypotonic lysis with NH4Cl. Cells were incubated first with anti-CD16/32 (eBioscience, San Diego, CA) to block Fc receptor binding to antibodies on macrophages, neutrophils and mast cells and stained with anti-CD3-FITC, anti-CD8-Pacific blue, anti-CD25-APC, anti-CD45R(B220)-Pe-Cy7 (eBioscience, San Diego, CA) and anti-CD4-PerCp (BD-Biosciences Pharmingen, San Diego, CA). Foxp3-positive cells were detected with PE anti-mouse/rat Foxp3 Staining Set, according to the manufacturer’s instruction (eBioscience, USA). Peripheral blood leukocytes were incubated with anti-CD11b-pacific blue (eBioscience, San Diego, CA) and anti-ly6G-PE (BD-Biosciences Pharmingen, San Diego, CA) to detect monocytes (CD11b+ly6G-) and granulocytes (CD11b+ly6G+). For measurement of inflammatory monocytes (CD11b+Ly6G-Ly6Chigh) cells were stained with anti-Ly6C-FITC (Miltenyi Biotec). Blood natural killer (NK) cells were stained by anti-CD49b(DX5)-APC (Miltenyi Biotec).
Donners M.M., Bai L., Lutgens S.P., Wijnands E., Johnson J., Schurgers L.J., Liu C.L., Daemen M.J., Cleutjens K.B., Shi G.P., Biessen E.A, & Heeneman S. (2016). Cathepsin K Deficiency Prevents the Aggravated Vascular Remodeling Response to Flow Cessation in ApoE-/- Mice. PLoS ONE, 11(9), e0162595.
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10

Spleen and MLN Cell Composition Analysis

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Six days after BMT, uninfected and Hpb-infected mice were sacrificed. The spleen and MLN were isolated for the analysis of cell composition. For surface staining, cells were suspended at 2×107 cells/ml in PBS with 2% FCS, and Fc receptors were blocked with a 2.4G2 mAb (Clone: 93, BioLegend). Antibodies for surface staining were: anti-CD3 FITC, anti-CD3 PE-Cy7 (Clone: 145–2C11), anti-CD4 PE-Cy7 (Clone: GK1.5; eBioscience), anti-H2b PE, anti-H2d PE, and anti-H2b APC (Clones: SF1–1.1, SF1–1.1.1, AF6.88.5; BD Biosciences). For intracellular Foxp3 staining, the Foxp3 staining buffer and anti-Foxp3 PE, Foxp3 PE-Cy7 or Foxp3 APC antibodies (Clone: FJK-16S; eBioscience) were used in accordance with the manufacturer’s instructions. For intracellular GATA3 staining, anti-GATA3 PE (Clone: L50–823; BD Biosciences) or isotype control IgG1, kappa was used.
Li Y., Guan X., Liu W., Chen H.L., Truscott J., Beyatli S., Metwali A., Weiner G.J., Zavazava N., Blumberg R.S., Urban JF J.r., Blazar B.R., Elliott D.E, & Ince M.N. (2018). Helminth-Induced Production of TGFβ and Suppression of Graft-Versus-Host Disease Is Dependent on Interleukin-4 Production by Host Cells. Journal of immunology (Baltimore, Md. : 1950), 201(10), 2910-2922.
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