Anti cd56 pe clone b159

Manufactured by BD

Sourced in United Kingdom, United States

Anti-CD56 PE (clone B159) is a monoclonal antibody conjugated with the fluorescent dye phycoerythrin (PE). The antibody binds specifically to the CD56 antigen, which is expressed on natural killer cells and a subset of T cells. This product can be used in flow cytometry applications to identify and enumerate these cell populations.

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Lab products found in correlation

Anti cd3 v450 clone ucht1, by BD (2 mentions) Anti cd4 apc h7 clone rpa t4, by BD (2 mentions) Anti cd8 pacific orange clone rpa t8, by BioLegend (2 mentions) Anti cd25 pe cy7 clone m a251, by BD (2 mentions) Anti cd127 pe cy5 clone ebiordr5, by Thermo Fisher Scientific (2 mentions) Anti cd19 apc clone hib19, by BD (2 mentions) Facscanto 2, by BD (2 mentions) Facsdiva software, by BD (2 mentions) Fortessa, by BD (1 mentions) Clone rpa t4, by BioLegend (1 mentions) Live dead fixable far red dead cell stain kit, by BD (1 mentions) Live dead fixable far red dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Lsrii instrument, by BD (1 mentions) Facsaria, by BD (1 mentions) Amcyan, by Thermo Fisher Scientific (1 mentions) Anti cd3 percp clone sk7, by BD (1 mentions) Anti ifnγ alexafluor 700, by BD (1 mentions) Anti cd107a apc clone h4a3, by BD (1 mentions) Brefeldin a, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

Close spelling variants of this product

Anti-CD56 PE-Cy7 (clone B159), Anti‐CD56 (clone B159; CD56 clone B159 Anti-CD56-PE CD56-PE Anti-CD56

5 protocols using anti cd56 pe clone b159

Comprehensive Immune Cell Profiling

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CD4⁺ T cells were defined as CD3+CD4⁺; CD8⁺ T cells were defined as CD3⁺CD8⁺; CD8⁺ naïve cells were defined as CD8⁺CD45RO-CD62L⁺; CD4 Tregs were defined as CD3⁺CD4⁺CD25^med-highCD127^low; CD4 conventional T cells (Tcons) were defined as CD3⁺CD4⁺CD25^low-negCD127^med-high; natural killer (NK) cells as CD56⁺CD3⁻; and B cells as CD19⁺. Aliquots of anti-coagulated whole blood (ethylenediaminetetraacetic acid [EDTA]) were incubated with fluorophore-conjugated monoclonal antibodies: anti-CD3 V450 (clone UCHT1, BD Biosciences), anti-CD4 APC-H7 (clone RPA-T4, BD Biosciences), anti-CD8 Pacific-Orange (clone RPA-T8, Biolegend), anti-CD25 PE-Cy7 (clone M-A251, BD Biosciences), anti-CD127 PE-Cy5 (clone eBioRDR5, eBioscience) for T-cell subsets; anti-CD56 PE (clone B159, BD Biosciences), anti-CD3 V450 (clone UCHT1, BD Biosciences) for NK/NKT cells, and anti-CD19 APC (clone HIB19, BD Biosciences) for B cells. RBC lysis with BD Pharm Lyse was performed either prior to or following incubation with conjugated antibodies. Flow cytometry analysis utilized FACSCanto II (BD Bioscience) or the Fortessa (BD Bioscience) and FACSDiva software (BD Bioscience). There was a change in the use of flow cytometry machines over the course of the study. Both flow cytometers were validated and results were comparable.

Koreth J., Kim H.T., Lange P.B., Poryanda S.J., Reynolds C.G., Rai S.C., Armand P., Cutler C.S., Ho V.T., Glotzbecker B., Yusuf R., Nikiforow S., Chen Y.B., Dey B., McMasters M., Ritz J., Blazar B.R., Soiffer R.J., Antin J.H, & Alyea EP I.I.I. (2018). Bortezomib-based immunosuppression after reduced-intensity conditioning hematopoietic stem cell transplantation: randomized phase II results. Haematologica, 103(3), 522-530.

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Comprehensive Immune Cell Profiling

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Immune reconstitution assays: CD4+ T cells were defined as CD3+CD4+; CD4+ naïve cells were defined as CD4+, CD45RO⁻; CD4+ memory cells were defined as CD4+CD45RO+; CD8+ T cells were defined as CD3+CD8+; CD8+ naïve cells were defined as CD8+CD45RO⁻CD62L+; CD8+ memory cells were defined as CD8+CD45RO+; CD8+ terminal effector cells were defined as CD8+CD45RO⁻CD62L⁻; CD4 regulatory T cells (Treg) were defined as CD3+CD4+CD25^med-highCD127^low; NK cells as CD56+CD3⁻; and B cells as CD19+. Fifty μl whole blood (15% EDTA) in 5 ml polystyrene round-bottom reaction tubes was incubated with fluorophore-conjugated monoclonal antibodies: anti-CD3 V450 (clone UCHT1, BD Biosciences), anti-CD4 APC-H7 (clone RPA-T4, BD Biosciences), anti-CD8 Pacific-Orange (clone RPA-T8, Biolegend), anti-CD25 PE-Cy7 (clone M-A251, BD Biosciences), anti-CD127 PE-Cy5 (clone eBioRDR5, eBioscience), anti-CD62L APC (clone DREG-56, BD Biosciences), CD45RO FITC (clone UCHL1, BD Biosciences) for T cell subsets; anti-CD56 PE (clone B159, BD Biosciences), anti-CD3 V450 (clone UCHT1, BD Biosciences) for NK/NKT cells; anti-CD19 APC (clone HIB19, BD Biosciences) for B cells. RBC lysis with 500 μl 1× BD Pharm Lyse followed. Immune reconstitution flow cytometry analysis utilized FACSCanto II (BD Bioscience) and FACSDiva software (BD Bioscience).

Koreth J., Kim H.T., Lange P.B., Bindra B., Reynolds C.G., Chammas M.J., Armand P., Cutler C.S., Ho V.T., Glotzbecker B., Nikiforow S., Ritz J., Blazar B.R., Soiffer R.J., Antin J.H, & Alyea EP I.I.I. (2015). A Bortezomib-based Regimen Offers Promising Survival and Graft-vs.-Host Disease Prophylaxis in Myeloablative HLA-Mismatched and Unrelated Donor Transplantation: A Phase II Trial. Biology of blood and marrow transplantation : journal of the American Society for Blood and Marrow Transplantation, 21(11), 1907-1913.

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PBMC Phenotyping Cocktails for T and NK Cells

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10⁵ total PBMC was stained in separate tubes with two phenotyping cocktails containing 2 μl of each antibody:
T cell antibody mix: anti-CD3—fluorescein isothiocyanate (FITC), clone UCHT1; anti-CD4—phycoerythrin (PE), clone RPA-T4; anti-CD8a-peridinin-chlorophyll protein—cyanine 5.5 (PerCP Cy5.5), clone RPA-8a (all BioLegend, London, UK), LIVE/DEAD Fixable Far Red Dead Cell Stain Kit (Thermo Fisher Scientific).
NK cell antibody mix: anti-CD3-FITC; anti-CD8-PerCP Cy5.5 (both as before); anti-CD56-PE, clone B159 (BD Pharmingen); LIVE/DEAD Fixable Far Red Dead Cell Stain Kit.

Houldcroft C.J., Jackson S.E., Lim E.Y., Sedikides G.X., Davies E.L., Atkinson C., McIntosh M., Remmerswaal E.B., Okecha G., Bemelman F.J., Stanton R.J., Reeves M, & Wills M.R. (2020). Assessing Anti-HCMV Cell Mediated Immune Responses in Transplant Recipients and Healthy Controls Using a Novel Functional Assay. Frontiers in Cellular and Infection Microbiology, 10, 275.

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Monoclonal Antibody Labeling and Transduction

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Monoclonal antibodies were purchased from e-biosciences (San Diego, CA): anti-human OKT3, anti-CD28 clone CD28-6; BD Biosciences (San Jose, CA): anti-CD16 FITC clone 3G8 BD, anti CD56 PE clone B159, anti-CD314/NKG2D PE or allophycocyanin, BD clone 1D11, isotype controls (IgG) as either FITC or PE conjugates; R&D systems (Minneapolis, MN): anti-human NKG2D clone 1 49810; Sigma Aldrich (St Louis, MO): anti-gamma Tubulin clone 4D11; Fisher Scientific (Pittsburg, PA) anti-HA HRP clone HA7, anti-Flag clone M2 HRP, anti-Myc HRP A5598; Santa Cruz Biotechnology (Santa Cruz, CA): anti-human NKG2D clone N-20, NKG2D peptide N-20, DAP10 peptide; Abcam (Cambridge, MA): anti-human NKG2D 5C6. All cell culture reagents and chemicals were purchased from Invitrogen (Grand Island, NY) and Sigma Aldrich, respectively, unless otherwise specified. COS-7 cells and HEK293T cells were cultured in complete DMEM media (cDMEM; 10% FCS; Invitrogen), 2 mM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin). The P815 murine mastocytoma cell line was purchased from the American Type Culture Collection (Manassas, VA) and cultured as recommended. Flow cytometry was performed on antibody-labeled or transduced cells by a BD LSR-II instrument (BD Biosciences). Transduced cells were cell-sorted with a FACSAria (BD Biosciences).

Karimi M.A., Aguilar O., Zou B., Bachmann M.H., Carlyle J.R., Baldwin C.L, & Kambayashi T. (2014). A truncated human NKG2D splice isoform negatively regulates NKG2D-mediated function. Journal of immunology (Baltimore, Md. : 1950), 193(6), 2764-2771.

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HIV-1 Antigen-Specific T-Cell Response Assay

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Whole blood samples were incubated with HIV-1 gag, pol and env peptide pools respectively at 37 °C for 5 h. Brefeldin-A (10 μg/ml, Sigma), GolgiStop (5 μg/ml, BD Biosciences) and anti-CD107a APC (clone H4A3, 20 μl/ml, BD Biosciences) were added immediately to the cell medium and incubated for 6 h. Samples incubated with phorbol-12-myristate-13-acetate (PMA) (1 mg/ml) plus ionomycin (1 mg/ml) were used as positive controls and samples incubated with DMSO (1 mg/ml, Sigma) were used as negative controls. Samples were surface-stained with live/dead dye Amcyan (Invitrogen, Carlsbad, CA, USA), anti-CD3-PerCP (clone SK7) and anti-CD56-PE (clone B159), and then lysed, permeabilized and intracellularly stained with anti-IFNγ Alexa Fluor700 (clone B27, BD Biosciences) and anti-CD107a APC (clone H4A3). All data were acquired on BD FACS Fortessa (BD Biosciences, San Jose, CA, USA) and analyzed by FlowJo software (Treestar, Ashland, OR, USA).

Fan X., Zhu L., Liang H., Xie Z., Huang X., Wang S, & Shen T. (2016). Antibody-dependent CD56+ T cell responses are functionally impaired in long-term HIV-1 infection. Retrovirology, 13, 76.

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