Anti humancd3 clone ucht1

Manufactured by BioLegend

Sourced in United States

Anti-human CD3 (clone UCHT1) is a monoclonal antibody that binds to the CD3 complex on human T cells. The CD3 complex is a key component of the T cell receptor and is involved in T cell activation.

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Lab products found in correlation

Cd8 clone rpa t8, by BioLegend (2 mentions) Cx3cr1 clone 2a9 1, by BioLegend (2 mentions) Software v10, by FlowJo (2 mentions) Lsrfortessa, by BD (2 mentions) Lymphocyte separation medium, by Corning (2 mentions) Anti human cd19 clone hib19, by BioLegend (1 mentions) Anti human cd27 clone o323, by BioLegend (1 mentions) Anti human cd38 clone hit2, by BioLegend (1 mentions) Anti human pd 1 clone eh12.2h7, by BioLegend (1 mentions) Live dead fixable dead cell stain, by Thermo Fisher Scientific (1 mentions) Anti human cd4 clone rpa t4, by BioLegend (1 mentions) Anti human cd21 cloneb ly4, by BD (1 mentions) Annexin 5, by BD (1 mentions) Human immunoglobulin g igg, by Merck Group (1 mentions) Anti human igg, by Merck Group (1 mentions) Cd45 clone hi30, by BD (1 mentions) Cd4 clone rpa t4, by BD (1 mentions) Cd19 clone hib19, by BioLegend (1 mentions) Percp, by BioLegend (1 mentions) Brilliant violet 421, by BD (1 mentions)

4 protocols using anti humancd3 clone ucht1

Multicolor Flow Cytometry Immunophenotyping

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The following fluorochrome-conjugated antibodies were used: anti-human
CD3 (clone UCHT1, 1:200), anti-human CD4 (clone RPA-T4, 1:200), anti-human CD19
(clone HIB19, 1:100), anti-human CD27 (clone O323, 1:100), anti-human CD38
(clone HIT2, 1:100), anti-human CD95 (clone DX2, 1:100), anti-human ICOS (clone
C398.4A, 1:100), anti-human CD40L (clone 24-31, 1:10), anti-human PD-1 (clone
EH12.2H7, 1:100), anti-human CXCR3 (clone G025H7, 1:200), anti-human CTLA4
(clone L3D10, 1:10), anti-human CD69 (clone FN50, 1:50) and anti-human
IL-7Rα (clone A019D5, 1:100) were all from BioLegend. Anti-human CXCR5
(clone RF8 B2, 1:200), anti-human Bcl-6 (clone K112-91, 1:20), anti-human IgD
(clone IA6-2, 1:50), anti-human Ki-67 (MOPC-21, 1:10) and anti-human CD21 (clone
B-ly4, 1:100) were from BD Biosciences and anti-human CD45RA (clone
2H4LDH11LDB9, 1:200) was from Beckman Coulter. LIVE/DEAD^®Fixable Dead Cell Stain from Invitrogen was used to gate on live cells as well
as Annexin V from BD Biosciences.

Cubas R., van Grevenynghe J., Wills S., Kardava L., Santich B.H., Buckner C.M., Muir R., Tardif V., Nichols C., Procopio F., He Z., Metcalf T., Ghneim K., Locci M., Ancuta P., Routy J.P., Trautmann L., Li Y., McDermott A.B., Koup R.A., Petrovas C., Migueles S.A., Connors M., Tomaras G.D., Moir S., Crotty S, & Haddad E.K. (2015). Reversible Reprogramming of Circulating Memory T Follicular Helper Cell Function during Chronic HIV Infection. The Journal of Immunology Author Choice, 195(12), 5625-5636.

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Isolation and Analysis of Human CX3CR1+ T Cells

Cited 1 time

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Using lymphocyte separation medium (Corning) density gradient centrifugation, we isolated peripheral blood mononuclear cells (PBMCs). Staining of CX3C chemokine receptor 1 (CX3CR1) in T cells was performed as described before [23 (link), 24 (link)]. In brief, fresh or cryopreserved PBMCs were incubated with Fc block with human immunoglobulin G (IgG) (Sigma) at 12 mg/mL for 20 min. Anti-human CD3 (clone UCHT1), CD8 (clone RPA-T8), and CX3CR1 (clone 2A9-1) antibodies were obtained from BioLegend, and anti-CD4 (clone RPA-T4) antibody was from BD Biosciences for flow cytometry. LSRFortessa (BD) was used for sample acquisition, and FlowJo software v10.1.5 (FlowJo LLC) was used for sample analysis.

Sarkar J., Cortes Gomez E., Oba T., Chen H., Dy G.K., Segal B.H., Ernstoff M.S, & Ito F. (2023). Fluctuations in Gut Microbiome Composition During Immune Checkpoint Inhibitor Therapy. World Journal of Oncology, 14(3), 178-187.

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Immune Cell Profiling of Peripheral Blood

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PB was obtained in EDTA-containing tubes. Peripheral blood mononuclear cells (PBMCs) were isolated using Lymphocyte Separation Medium (Corning) density gradient centrifugation. Fresh or cryopreserved PBMCs were incubated with anti-human IgG (Sigma). These antibodies were used for flow cytometry or cell sorting: anti-human CD3 (clone UCHT1; BioLegend), CD4 (clone RPA-T4; BD Biosciences), CD8 (clone RPA-T8; BioLegend), CD45 (clone HI30; BD Biosciences), CD56 (cone HCD56: BioLegend), CD19 (clone HIB19 BioLegend), and CX3CR1 (clone 2A9–1; BioLegend) antibodies. Samples were acquired using LSRFortessa (BD Biosciences), and data were analyzed with FlowJo software v10.1.5 (FlowJo LLC).

Abdelfatah E., Long M.D., Kajihara R., Oba T., Yamauchi T., Chen H., Sarkar J., Attwood K., Matsuzaki J., Segal B.H., Dy G.K, & Ito F. (2023). Predictive and Prognostic Implications of Circulating CX3CR1+ CD8+ T Cells in Non–Small Cell Lung Cancer Patients Treated with Chemo-Immunotherapy. Cancer Research Communications, 3(3), 510-520.

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Multi-color Flow Cytometry Analysis

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Cells were stained with fluorochrome conjugated monoclonal antibodies: PerCP or APC (from Biolegend, San Diego, California, USA) or Brilliant Violet 421 (from BD Biosciences, Franklin Lakes, New Jersey, USA), antihuman CD3, clone UCHT1, together with PE conjugated anti-human TCR γδ, clone B1 (from Biolegend) or APC anti-human TCR γδ, clone B1 (BD Biosciences) and APC conjugated anti-human CD27, clone L128, and PE-cy7 conjugated anti-human CD45RA, clone L48 (both from BD Biosciences) for 30 min at 4°C. The cells were then stained with 7-aminoactinomycin D (7-AAD, BD Biosciences Pharmingen) for 10 min at 4°C for visualization of viable 7-AAD negative cells (7-AAD^-) and analyzed on a BD FACS Sort using the CellQuest software (BD Biosciences) or Beckman Coulter Gallios. Further analysis was performed using the software FlowJo 9.4.10 (Tree Star Inc, Ashland, Oregon, USA). APC conjugated anti-human IFN-γ clone B27 (Biolegend) was used for intracellular staining.

Laurent A.J., Bindslev N., Johansson B, & Berg L. (2014). Synergistic Effects of Ethanol and Isopentenyl Pyrophosphate on Expansion of γδ T Cells in Synovial Fluid from Patients with Arthritis. PLoS ONE, 9(8), e103683.

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