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Cd3 fitc ucht1

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CD3-FITC (UCHT1) is a fluorescently-labeled antibody that binds to the CD3 antigen on the surface of T cells. It is commonly used in flow cytometry applications to identify and enumerate T cell populations.

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Lab products found in correlation

7 aminoactinomycin d, by BD (1 mentions) 7 aad, by BD (1 mentions) Cd4 apc sk3, by BD (1 mentions) Facsaria, by BD (1 mentions) 7 aad, by Thermo Fisher Scientific (1 mentions) Dylight 650, by Thermo Fisher Scientific (1 mentions) Imagestream x mark 2 imaging flow cytometer, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Cytofix perm kit, by BD (1 mentions)

Spelling variants of this product

CD3-FITC (243 citations) CD3-FITC (clone UCHT1, (7 citations) Anti-CD3 FITC (clone: UCHT1; (3 citations) Anti-CD3 FITC (UCHT1) (2 citations)

4 protocols using cd3 fitc ucht1

1

Immunosuppression Monitoring in Upper Extremity Transplant

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A male patient underwent unilateral upper extremity transplantation at MGH in October 2012 as previously reported27 (link). Immunosuppression was induced with rabbit antithymocyte globulin, and maintained with Tacrolimus, Mycophenolate Mofetil and Prednisolone. Protocol biopsies were obtained at 6 monthly intervals post-transplant with the consent of the patient and in accordance with the Institutional Review Board protocol. At 12- and 18-month points, samples were processed for flow cytometry as described above, stained with CD3-FITC (UCHT1, BD Biosciences, USA), and HLA A1/B8 (One Lambda, USA) conjugated to Streptavidin (0.1ug/ml, BD Biosciences, USA) and acquired on LSRII (Becton Dickinson, USA). Analysis was performed using FlowJo (TreeStar Inc., USA). Samples were also processed by routine histological methods and assessed by board certified pathologist.
Leonard D.A., Powell H.R., Defazio M.W., Shanmugarajah K., Mastroianni M., Rosales I., Farkash E.A., Colvin R.B., Randolph M.A., Sachs D.H., Kurtz J.M, & Cetrulo CL J.r. (2020). Cutaneous leukocyte lineages in tolerant large animal and immunosuppressed clinical vascularized composite allograft recipients. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(2), 582-592.
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2

PBMC Proliferation Assay with Flow Cytometry

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As described previously (3 (link)), proliferation assays consisted of three replicate wells per condition containing ~0.3 million in 225 μl per well of CTV-labelled PBMC and 25 μl test solution. After 8 days cells were stained with CD3-FITC (UCHT1), CD4-APC (SK3), and 7-Amino-Actinomycin D to discriminate dead cells (7-AAD; all from BD Biosciences). Cells were analysed on a BD FACSVerse cytometer and flow cytometry data was analysed by FlowJo software (version 10; FlowJo, LLC).
Hardy M.Y., Goel G., Russell A.K., Chen Yi Mei S.L., Brown G.J., Wang S., Szymczak E., Zhang R., Goldstein K.E., Neff K.M., Williams L.J., Truitt K.E., Dzuris J.L., Tye-Din J.A, & Anderson R.P. (2021). A Sensitive Whole Blood Assay Detects Antigen-Stimulated Cytokine Release From CD4+ T Cells and Facilitates Immunomonitoring in a Phase 2 Clinical Trial of Nexvax2 in Coeliac Disease. Frontiers in Immunology, 12, 661622.
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3

Isolation and Characterization of MuSK-Specific Memory B Cells

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MuSK-binding memory B cells were isolated using a fluorescence-activated cell sorter (FACSaria; BD Biosciences, San Jose, CA) from cryopreserved peripheral blood mononuclear cells (using mouse anti-human monoclonals CD19-BV421 HIB19, CD20-AF700 2H7, CD27-APCHy7 M-T271 all from BD Biosciences; in 0.1 % BSA, 2 mM EDTA/Dulbecco's PBS). To remove dead cells and non–B cells, a dump channel was included (7-AAD,00-6993-50, CD3/FITC UCHT1 BD and CD14 (labeled with 7-AAD from Thermo Fisher, Waltham, MA; CD3/FITC UCHT1 and CD14/FITC M5E2 from BD Biosciences; and CD56/FITC HCD56 from BioLegend, San Diego, CA). Antigen-specific cells were isolated using recombinant MuSK produced in Escherichia coli6 (link) labeled with R-PE (AS-72113, AnaSpec) and MuSK produced in yeast tagged with DyLight 650 (a kind gift of Konstantinos Lazaridis and Socrates Tzartos, Thermo Fisher). Single sorted cells were cultured on irradiated CD40L cells (a kind gift from Kees van Kooten) in a 96-well plate in complex RPMI medium.12 (link) After 2 weeks, the medium was tested in duplicate for MuSK antibody production using the MuSK ELISA as described previously.6 (link)
Huijbers M.G., Vergoossen D.L., Fillié-Grijpma Y.E., van Es I.E., Koning M.T., Slot L.M., Veelken H., Plomp J.J., van der Maarel S.M, & Verschuuren J.J. (2019). MuSK myasthenia gravis monoclonal antibodies: Valency dictates pathogenicity. Neurology® Neuroimmunology & Neuroinflammation, 6(3), e547.
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4

Quantifying Nuclear Localization of IRF5

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PBMCs or enriched pDCs were stained with the following cell surface markers: CD3-FITC (UCHT1, BD Biosciences), CD14-AF488 (M5E2, BD Biosciences), CD19-FITC (HIB19, BD Biosciences), CD56-AF488 (B159, BD Biosciences), CD11c-FITC, HLA-DR-PE (L243, Biolegend), BCDA2-PE (AC144, Miltenyi), FITC-conjugated lineage cocktail panel 1 (BD Biosciences). Cells were fixed and permeabilized using the Cytofix/perm kit (BD), and stained with IRF5-AF647 antibody (EPR6094, Abcam) and DAPI (Life Technologies). Stained cells were acquired on an ImageStreamX Mark II imaging flow cytometer at 60x magnification (Amnis/EMD Millipore). At least 10,000 cells per condition were analyzed. Data were compensated and analyzed using the IDEAS software. Following standard analysis procedures(26 (link)), single focused cells were analyzed using Nuclear Localization Wizard to calculate similarity score (SS) between IRF5 and DAPI, which quantifies the correlation of pixel values of the two stains on a per cell basis. Representative images of cells and/or SS plotted on histograms are displayed.
Chow K.T., Wilkins C., Narita M., Green R., Knoll M., Loo Y.M, & Gale M J.r. (2018). Differential and overlapping immune programs regulated by IRF3 and IRF5 in plasmacytoid dendritic cells. Journal of immunology (Baltimore, Md. : 1950), 201(10), 3036-3050.
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