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Cd3 v500 clone sp34 2

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The CD3 V500 (clone SP34-2) is a laboratory equipment product manufactured by BD. It is a fluorochrome-conjugated monoclonal antibody that binds to the CD3 antigen, which is expressed on the surface of T cells. The core function of this product is to facilitate the identification and enumeration of T cells in various biological samples through flow cytometry analysis.

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Lab products found in correlation

Cd8 af 647 clone rpa t8, by BD (2 mentions) Cd20 apc h7 clone 2h7, by BD (2 mentions) Cd14 pe cy7 clone m5e2, by BD (2 mentions) Vacutainer cpt tube, by BD (2 mentions) Cd150 bv 421, by BD (1 mentions) Macsquant 10 flow cytometer, by Miltenyi Biotec (1 mentions) Cd38 pe cy7, by BD (1 mentions) Ccr5 pe clone 3a9, by BD (1 mentions) Cd4 percp cy5.5 clone l200, by BD (1 mentions) Methanol free formaldehyde, by Polysciences (1 mentions) Cyan adp, by Beckman Coulter (1 mentions) Cd38 pe cy7 clone hb7, by BD (1 mentions) Kaluza 2, by Beckman Coulter (1 mentions) Macsquant 10, by Miltenyi Biotec (1 mentions) Macsquant10 analyzer, by Miltenyi Biotec (1 mentions) Ifnγ apc clone b27, by BD (1 mentions) Cd4 fitc clone rpa t4, by BD (1 mentions) Cd8 v450 clone rpa t8, by BD (1 mentions) Cytofix, by BD (1 mentions)

4 protocols using cd3 v500 clone sp34 2

1

PBMC Isolation and Immunophenotyping

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Whole blood was collected on EDTA, then transferred into BD vacutainer CPT tubes (after removal of anticoagulant solution from CPT tubes) and spun at 2500g for 20 min. The PBMCs were collected and one-tenth were used to isolate RNA. The remaining cells were surface stained on ice using three different panels, including Panel A: CD150 BV-421 (clone A12, BD), CD8 AF 647 (clone RPA-T8), CD20 APC-H7 (clone 2H7, BD), CD3 V500 (clone SP34–2, BD), CD14 Pe-cy7 (clone M5E2, BD); Panel B: CD150 BV421, CD3 V500, CCR7 Pe-Cy7 (clone G043H7, Biolegend), CD8 AF647, CD45RA APC-H7 (clone 5H9, BD); and Panel C: CD150 BV421, IgD BV510 (clone IA6–2, BD), CD38 Pe-Cy7 (clone HB7, BD), CD27 AF647 (clone O323, Biolegend), CD20 APC-H7. Cells were acquired on a MACSQuant®10 flow cytometer (Miltenyi).
Reynard O., Gonzalez C., Dumont C., Iampietro M., Ferren M., Le Guellec S., Laurie L., Mathieu C., Carpentier G., Roseau G., Bovier F.T., Zhu Y., Le Pennec D., Montharu J., Addetia A., Greninger A.L., Alabi C.A., Moscona A., Vecellio L., Porotto M, & Horvat B. (2022). Nebulized fusion inhibitory peptide protects cynomolgus macaques from measles virus infection. Research Square.
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2

PBMC Phenotyping by Flow Cytometry

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PBMC were stained with various combinations of the following monoclonal antibodies: CD3-V500 (clone SP34.2, BD-Biosciences), CD4-PerCp-Cy5.5 (clone L200, BD-Biosciences) or CD4-APC, clone 13B8.2, Beckman-Coulter), CD8-FITC (clone 3B5, Invitrogen, ThermoFisher), CCR5-PE (clone 3A9, BD-Biosciences). After 30 min of incubation at 4°C, cells were washed with cold PBS then fixed in PBS containing 1.6% methanol-free formaldehyde (Polysciences). Data was collected on a three-laser CyAn ADP (Beckman-Coulter) and analyzed on FlowJo version 10 software.
Obregon-Perko V., Hodara V.L., Parodi L.M, & Giavedoni L.D. (2018). Baboon CD8 T cells suppress SIVmac infection in CD4 T cells through contact-dependent production of MIP-1α, MIP-1β, and RANTES. Cytokine, 111, 408-419.
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3

PBMC Isolation and Multiparameter Flow Cytometry

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Whole blood was collected on EDTA, then transferred into BD vacutainer CPT tubes (after removal of anticoagulant solution from CPT tubes) and spun at 2500 g for 20 min. The PBMCs were collected and one-tenth were used to isolate RNA. The remaining cells were surface stained on ice using three different panels, including Panel A: CD150 BV-421 (clone A12, BD), CD8 AF 647 (clone RPA-T8), CD20 APC-H7 (clone 2H7, BD), CD3 V500 (clone SP34-2, BD), CD14 Pe-cy7 (clone M5E2, BD); Panel B: CD150 BV421, CD3 V500, CCR7 Pe-Cy7 (clone G043H7, Biolegend), CD8 AF647, CD45RA APC-H7 (clone 5H9, BD); and Panel C: CD150 BV421, IgD BV510 (clone IA6-2, BD), CD38 Pe-Cy7 (clone HB7, BD), CD27 AF647 (clone O323, Biolegend), CD20 APC-H7, using 4 µl of each Ab per sample. Cells were acquired on a MACSQuant®10 flow cytometer (Miltenyi) and analyzed by FlowJo10.8 (Becton Dickinson) and Kaluza 2.1 (Beckman Coulter). Gating strategy is presented in supplementary fig. S8.
Reynard O., Gonzalez C., Dumont C., Iampietro M., Ferren M., Le Guellec S., Laurie L., Mathieu C., Carpentier G., Roseau G., Bovier F.T., Zhu Y., Le Pennec D., Montharu J., Addetia A., Greninger A.L., Alabi C.A., Brisebard E., Moscona A., Vecellio L., Porotto M, & Horvat B. (2022). Nebulized fusion inhibitory peptide protects cynomolgus macaques from measles virus infection. Nature Communications, 13, 6439.
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4

Multiparametric Analysis of T Cell Polyfunctionality

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Cryopreserved stimulated whole blood samples were slowly thawed in a 37°C water bath, followed by fixation and permeabilization using Cytofix (BD Biosciences, 554722) and Cytoperm/Wash (BD Biosciences, 554723) according to the manufacturer’s instructions. Cells were stained with the following fluorochrome conjugated antibodies to assess polyfunctionality and memory: CD8 V450 (clone RPA-T8, Cat 560347), CD3 V500 (clone SP34-2, Cat 560770), CD4 FITC (clone RPA-T4, Cat 555346 and clone M-T477, Cat 556615), CD45RA-PE (clone HI100, Cat 555489) and IFN-γ APC (clone B27, Cat 554702) all from BD Biosciences as well as IL-2 PE (clone N7.48A, Beckman, Cat IM2718U) and TNF PCP-Cy5.5 (clone MAb11, eBioscience, Cat 45-7349-42). Acquisition of a minimum of 100,000 cells per sample was performed on a MACSQuant 10 analyzer (Miltenyi Biotec). Single-stained controls using mouse Ig/ κ compensation beads (BD Biosciences, 552843) were used to calculate compensation. Flow cytometry data was analyzed using FlowJo software v10.5.3 software (BD Life Sciences) [40 ]. Boolean combination gating was performed to assess polyfunctionality of T cells. The gating strategy for flow cytometry analysis of T cell-specific (CD4+ or CD8+) cytokine expression is presented in S1 Fig.
Inthawong M., Pinthong N., Thaiprakhong A., Wangrangsimakul T., Sunyakumthorn P., Hill J., Sonthayanon P., Paris D.H., Dunachie S.J, & Kronsteiner B. (2023). A whole blood intracellular cytokine assay optimised for field site studies demonstrates polyfunctionality of CD4+ T cells in acute scrub typhus. PLOS Neglected Tropical Diseases, 17(3), e0010905.
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