Cd3 apcefluor780 ucht1

Manufactured by Thermo Fisher Scientific

CD3-APCefluor780 (UCHT1) is a fluorochrome-conjugated monoclonal antibody that binds to the CD3 epsilon chain on the surface of T cells. It is a tool used for the identification and enumeration of T cells in flow cytometry applications.

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Lab products found in correlation

Lsrii flow cytometer, by BD (2 mentions) Cd4 qdot655 s3, by Thermo Fisher Scientific (2 mentions) Cd8 qdot 605 3b5, by Thermo Fisher Scientific (2 mentions) Hla dr pe l243, by BD (2 mentions) Facsdiva software version 6, by BD (1 mentions) Cd45 af700, by BioLegend (1 mentions) Sytox blue, by Thermo Fisher Scientific (1 mentions) Cd4 apc sk3, by Thermo Fisher Scientific (1 mentions)

3 protocols using cd3 apcefluor780 ucht1

Multiparameter Flow Cytometry of T Cell Subsets

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Methods have been previously reported [20 (link)]. Briefly, cryopreserved PBMCs were thawed and stained with surface antibodies against CD3-APCefluor780 (UCHT1; eBiosciences, San Diego, CA); CD4-Qdot 655 (S3.5) and CD8-Qdot 605 (3B5) (both from Life Technologies, Eugene, OR); CXCR3-APC-CD183 (1C6), CCR4 BV421-CD194 (1G1), CCR5-PE-Cy7-CD195 (2D7), CCR7-Alexa Fluor700-CD197 (150503), CD38-FITC (HB7) and HLA-DR-PE (L243) (all from BD Biosciences, San Jose, CA). Cells were then incubated for 30 minutes and centrifuged at 1200 rpm for 10 minutes. PBMCs were fixed with 2% formalin before analysis. Stained cells were acquired using an LSR II flow cytometer (BD Immunocytometry Systems, San Jose, CA). Using 10-color flow cytometric analysis, we investigated T cell phenotypes classified by the markers listed above. Data were analyzed using FlowJo software (v9.8.5, TreeStar, Ashland, OR). Lymphocytes were analyzed based on forward scatter (FSC) and side scatter (SSC) profiles. A single cell population was selected with SSC-A vs. SSC-H and then with FSC-A vs. FSC-H before gating for other markers. Gates were set based on Fluorescence Minus One (FMO) control.

Ogendi B.M., Bakshi R.K., Gupta K., Kapil R., Brown L.T., Jordan S.J., Sabbaj S., Press C.G., Lee J.Y, & Geisler W.M. (2017). T cell phenotypes in women with Chlamydia trachomatis infection and influence of treatment on phenotype distributions. Microbes and infection, 20(3), 176-184.

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Phenotypic Characterization of Lymphocytes

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Peripheral blood mononuclear cells (PBMCs) were isolated using density centrifugation. PBMCs and aliquots of donor lymphocytes were analyzed immediately or cryopreserved. Thawed PBMCs were stained in MACS buffer on ice for 30 min using pre-determined concentrations of the following antibodies: CD14-Pacific Blue (M5E2), TCRα/β-AF488 (IP26), CD56-PE (HCD56), CD8-PerCP-Cy5.5 (SK1), and CD45-AF700 (HI30) from Biolegend; CD19-PE-Cy7 (HIB19), CD4-APC (SK3), and CD3-APC-eFluor780 (UCHT1) from eBioscience. After washing in MACS buffer, and addition of Sytox Blue (Invitrogen) for dead cell discrimination, cells were acquired on an LSRII (BD) flow cytometer (Beckman Coulter). Doublets, CD14+ monocytes and dead cells were excluded before a minimum of 30,000 CD45+ lymphocyte events were recorded. Data were analyzed using FacsDIVA software version 6.1.3 (BD). Lymphocyte counts, from routine clinical analysis of whole blood, were used to calculate lymphocyte subset frequency.

Burns D.M., Ryan G.B., Harvey C.M., Nagy E., Hughes S., Murray P.G., Russell N.H., Fox C.P, & Long H.M. (2019). Non-uniform in vivo Expansion of Epstein-Barr Virus-Specific T-Cells Following Donor Lymphocyte Infusion for Post-transplant Lymphoproliferative Disease. Frontiers in Immunology, 10, 2489.

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Multiparameter Flow Cytometry for PBMC and MMC Characterization

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PBMCs and MMCs were stained with fluorescent antibodies against cell surface markers and analyzed using 10-color flow cytometry by previously reported methodology [3 (link)]. Briefly, PBMCs and MMCs were labeled with LIVE/DEAD fluorescent reactive dye (Invitrogen, Carlsbad, CA) and stained with surface antibodies against CD3-APCefluor780 (UCHT1; eBiosciences, San Diego, CA); CD4-Qdot 655 (S3.5) and CD8-Qdot 605 (3B5) (both from Life Technologies, Eugene, OR); CXCR3-APC-CD183 (1C6), CCR4 BV421-CD194 (1G1), CCR5-PE-Cy7-CD195 (2D7), CCR7-Alexa Fluor700-CD197 (150503), CD38-FITC (HB7) and HLA-DR-PE (L243) (all antibodies were purchased from BD Biosciences, San Jose, CA). Cells were then incubated for 30 minutes and centrifuged at 1200 rpm for 10 minutes. Both PBMCs and MMCs were fixed with 2% formalin before analysis. Using an LSR II flow cytometer (BD Immunocytometry Systems, San Jose, CA), CD3⁺ events were acquired from each sample. Data were analyzed using FlowJo software (v9.8.5, TreeStar, Ashland, OR). Lymphocytes analysis was based on forward and side scatter profiles after exclusion of dead cells (Fig 1). Gates were either set using negative sample or Fluorescence Minus One (FMO) control and were then applied to all samples from each participant.

Ogendi B.M., Bakshi R.K., Sabbaj S., Brown L., Lee J.Y., Kapil R, & Geisler W.M. (2017). Distinct peripheral versus mucosal T cell phenotypes in chlamydia-infected women. American journal of reproductive immunology (New York, N.Y. : 1989), 78(6), 10.1111/aji.12768.

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