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Cd27 bv510

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The CD27 BV510 is a fluorophore-conjugated antibody that binds to the CD27 antigen, which is a member of the tumor necrosis factor receptor superfamily. It is used for the detection and quantification of CD27-expressing cells in flow cytometry applications.

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Lab products found in correlation

Cd19 pe cy7, by BD (2 mentions) Facs aria fusion cytometer, by BD (1 mentions) Cd14 percpcy5, by Thermo Fisher Scientific (1 mentions) Cd19 pe cy7, by BioLegend (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Igm af488, by BioLegend (1 mentions) Streptactin apc, by IBA Lifesciences (1 mentions) Strep tactin pe, by IBA Lifesciences (1 mentions) Cd8 percp cy5, by BioLegend (1 mentions) Anti human cd3 percp cy5, by BioLegend (1 mentions) Cd20 pe cy7, by BioLegend (1 mentions) Cd16 percp cy5, by BioLegend (1 mentions) Igd bv421, by BioLegend (1 mentions) Macs human b cell isolation kit, by Miltenyi Biotec (1 mentions) Cxcr5 percp cy5, by BioLegend (1 mentions) Cd4 apc vio770, by Miltenyi Biotec (1 mentions) Cd45ra pe vio770, by Miltenyi Biotec (1 mentions) Ccr6 apc, by Miltenyi Biotec (1 mentions) Cxcr3 viobright fitc cxcr5 percp cy5, by Miltenyi Biotec (1 mentions) Ccr4 pe, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

Anti-CD27-BV510 (2 citations)

6 protocols using cd27 bv510

1

Tetramer-based Identification and Sorting of B Cells

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B cells were eluted from PBMCs using a MACS Human B Cell isolation kit (Miltenyi Biotec). B cells were stained with rGP38 (IbAr10200) that had been tetramerized at 25 nM using Streptactin-PE (IBA Lifesciences) and Streptactin-APC (IBA Lifesciences). B cells were simultaneously stained with rGP38-Streptactin-PE and rGP38-Streptactin-APC tetramers for 1 hour on ice. Cells were washed twice in buffer (PBS, FBS, EDTA). Next, B cells were stained with a panel of antibodies. Donor 1 PBMCs were stained with a cocktail of anti-human CD3 PerCP-Cy5.5 (Biolegend), CD8 PerCP-Cy5.5 (Biolegend), CD14 PerCP-Cy5.5 (Invitrogen), CD16 PerCP-Cy5.5 (Biolegend), propidium iodide (PI) (Invitrogen), CD19 PE-Cy7 (Biolegend), CD27 BV510 (BD Biosciences), IgM BV711 (BD Biosciences), IgD BV421 (Biolegend), IgG BV605 (BD Biosciences), and IgA AF488 (Abcam). Donor 5 and 6 PBMCs were stained with a cocktail of anti-human CD3 PerCP-Cy5.5 (Biolegend), CD8 PerCP-Cy5.5 (Biolegend), CD14 PerCP-Cy5.5 (Invitrogen), CD16 PerCP-Cy5.5 (Biolegend), PI (Invitrogen), CD19 PE-Cy7 (Biolegend), CD20 PE-Cy7 (Biolegend), CD27 BV510 (BD Biosciences), IgM AF488 (Biolegend), and IgD BV421 (Biolegend). B cells were washed twice in buffer and run on a FACS Aria Fusion Cytometer (BD Biosciences). B cells were sorted into Super Script III reaction buffer (ThermoFisher Scientific) in 96-well Costar plates and frozen at −80 °C.
Shin O.S., Monticelli S.R., Hjorth C.K., Hornet V., Doyle M., Abelson D., Kuehne A.I., Wang A., Bakken R.R., Mishra A., Middlecamp M., Champney E., Stuart L., Maurer D.P., Li J., Berrigan J., Barajas J., Balinandi S., Lutwama J.J., Lobel L., Zeitlin L., Walker L.M., Dye J.M., Chandran K., Herbert A.S., Pauli N.T, & McLellan J.S. (2024). Crimean-Congo Hemorrhagic Fever Survivors Elicit Protective Non-Neutralizing Antibodies that Target 11 Overlapping Regions on Viral Glycoprotein GP38. bioRxiv.
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2

Multicolor Flow Cytometry Analysis of Lymphocyte Subsets

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Multicolour flow cytometric analysis was performed using the following fluorochrome-labelled anti-human monoclonal antibodies for measuring lymphocyte Th and Tfh subsets (CD4-APC-Vio770, CD45RA-PE-Vio770, CCR6-APC, CXCR3-VioBright FITC CXCR5 PerCP-Cy5.5, CCR4-PE) all from Miltenyi biotech. Tregs and Tfr were detected using BD Human Regulatory T Cell Cocktail (BD, San Jose, CA, USA), including CD4-FITC (clone SK3), CD25-PECy7 (clone 2A3), CD127 Alexa Fluor® 647 and adding CXCR5 PerCP-Cy5 and CD8 horizon450 (the last two from Biolegend). Bregs were detected using the following fluorochrome-labelled anti-human monoclonal antibodies: CD38-FITC CD1d-PE, CD19-PE-Cy7, CD5-PerCP-CY5.5, CD24-APC-H7 and CD27- BV510, all from BD Biosciences (San Jose, CA, USA).
Bergantini L., d’Alessandro M., Cameli P., Pianigiani T., Fanetti M., Sestini P, & Bargagli E. (2021). Follicular T Helper and Breg Cell Balance in Severe Allergic Asthma Before and After Omalizumab Therapy. Molecular Diagnosis & Therapy, 25(5), 593-605.
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3

Comprehensive Immunophenotyping of Bone Marrow

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Characterization of human samples was performed using the EuroFlow lyse-wash-and-stain using a standard sample preparation protocol adjusted to 106 BM-derived nucleated cells, together with the eight-color combination of the monoclonal antibodies CD138-BV421, CD27-BV510, CD38-FITC, CD56-PE, CD45-PerCPCy5.5, CD19-PECy7, CD117-APC and CD81-APCH7 (BD Biosciences)67 (link). Data acquisition was performed in a FACS CantoII flow cytometer (BD Biosciences). Samples were analyzed using the Infinicyt software (Cytognos SL) and the semiautomated pipeline ‘FlowCT’, based on the analysis of multiple files by automated cell clustering68 (link). Cell sorting was performed in a FACS Aria sorter instrument. Classification of BM samples according to immune cell infiltration was calculated similar to that in the mouse samples. The maximum percentages of T cells and NK cells present in the BM from healthy control individuals (cutoff, 20%) were used to divide patients with MM into cases with low or high number of immune-infiltrating cells.
Larrayoz M., Garcia-Barchino M.J., Celay J., Etxebeste A., Jimenez M., Perez C., Ordoñez R., Cobaleda C., Botta C., Fresquet V., Roa S., Goicoechea I., Maia C., Lasaga M., Chesi M., Bergsagel P.L., Larrayoz M.J., Calasanz M.J., Campos-Sanchez E., Martinez-Cano J., Panizo C., Rodriguez-Otero P., Vicent S., Roncador G., Gonzalez P., Takahashi S., Katz S.G., Walensky L.D., Ruppert S.M., Lasater E.A., Amann M., Lozano T., Llopiz D., Sarobe P., Lasarte J.J., Planell N., Gomez-Cabrero D., Kudryashova O., Kurilovich A., Revuelta M.V., Cerchietti L., Agirre X., San Miguel J., Paiva B., Prosper F, & Martinez-Climent J.A. (2023). Preclinical models for prediction of immunotherapy outcomes and immune evasion mechanisms in genetically heterogeneous multiple myeloma. Nature Medicine, 29(3), 632-645.
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4

Flow Cytometric Analysis of Surface Markers

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To analyse expression of surface markers, MCs were incubated with fluorochrome‐labeled monoclonal antibodies for 30 min in the dark on ice and fluorescence intensity was measured using a LSR II flow cytometer (BD Biosciences). CD3 V500, CD3 Alexa Fluor 647, CD4 PerCP‐Cy5.5, CD8 APC‐H7, CD14 V500, CD14 Pe‐Cy7, CD27 BV510, CD43 FITC, CD45RA PeCy‐7, CD56 BV510 (BD Biosciences) and Siglec‐1 Alexa Fluor 647 (Sanbio) were used. Relative mean intensity fluorescence (MFI) of CD43 was calculated as follows: MFI stained/MFI unstained. GFP fluorescence was used to determine replication of (GFP)‐labeled RSV.
Jans J., Unger W.W., Vissers M., Ahout I.M., Schreurs I., Wickenhagen A., de Groot R., de Jonge M.I, & Ferwerda G. (2018). Siglec‐1 inhibits RSV‐induced interferon gamma production by adult T cells in contrast to newborn T cells. European Journal of Immunology, 48(4), 621-631.
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5

Comprehensive Immune Cell Profiling

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The following mAbs were used to detect surface markers: anti-CD3 APC-Cy7 (clone: UCHT1), CD14 APC-Cy7 (HCD14), CD16 BV510 (3G8), CD19 APC-Cy7 (HIB19), CD25 BV421 (BC96), CD69 BV421 (FN50), CD56 PE-Cy7 (HCD56), CD4 FITC (RPAT4) and CD62L (DREG-56), all IgG1 isotypes from Biolegend. CD57 VioBlue (TB03. isotype IgM) CD159C PE (REA 205, isotype IgG1), CD279 PE (PD1.3.1.3. isotype: IgG2B), CD8 Vioblue (REA734 isotype IgG1) and CD45RA PE-Vio770 (REA 562. isotype IgG1) were from Miltenyi Biotech Bergisch, Gladbach, Germany, CD158a FITC (HP-3E4 isotype IgM), CD158b FITC (CH-L. isotype IgG2b) and CD27 BV510 (L128. isotype IgG1) were from BD Biosciences. CA. USA. CD159a APC (Z199. isotype IgG2b) and CD336 PE (Z231. Isotype IgG1) were from Beckman Coulter, (Beckman Coulter. 250 S. Kraemer Blvd, Brea, CA 92821, USA).
Bergantini L., d’Alessandro M., Cameli P., Cavallaro D., Gangi S., Cekorja B., Sestini P, & Bargagli E. (2021). NK and T Cell Immunological Signatures in Hospitalized Patients with COVID-19. Cells, 10(11), 3182.
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6

Phenotypic Analysis of Cryopreserved PBMCs

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Cryopreserved PBMCs were thawed at 37°C, washed in RPMI/60% and RPMI/20% of fetal bovine serum (FBS), and incubated for 1 h at 37°C in RPMI/10%FBS. PBMCs were then stained with the Fixable Viability Stain-FVS780r (APC-H7 detect, BD Biosciences) for 15 mins. After PBMCs washing in PBS/1%FBS, cells were incubated in a U-bottom 96-well plate at a density of 1.5 million/well and stained with selected 14-color panel including: anti-CD19-AF700 (Clone HIB19), anti-IgD-Pe-Cy7 (Clone IA6-2), anti-IgM-BB515 (Clone G-20-127), polyclonal goat anti-IgA-Dylight®649 (from Jackson Immunoresearch), CD10-BV650 (Clone H10a), CD21-PE-CF594 (Clone B-LY4), CD27-BV510 (Clone L128), CD38-BV786 (Clone HIT2), CD45-RB-PE (Clone MT4), CD86-PerCP-Cy5.5 (Clone 2331), CD279 (PD-1)-BV421 (Clone EH12.1), all from BD Biosciences unless indicated, for 15 mins. After washing twice in PBS/1% FBS, cells were fixed in PBS/1% formaldehyde, acquired in a BD LSRFortessa (BD Biosciences) cytometer using a plate HTS loader (BD Biosciences) and analyzed with FlowJo software (Tree Star). Gating strategy is described in Supplementary Figure 1. Lymphocyte gate was defined manually by morphological parameters excluding non-viable cells. B cells were identified as CD19+ CD21+/− cells and gated according to the expression of different markers to identify B-cell maturation stages as described in Supplementary Figure 1.
Jiménez M., Pastor L., Urrea V., Rodríguez de la Concepción M.L., Parker E., Fuente-Soro L., Jairoce C., Mandomando I., Carrillo J., Naniche D, & Blanco J. (2021). A Longitudinal Analysis Reveals Early Activation and Late Alterations in B Cells During Primary HIV Infection in Mozambican Adults. Frontiers in Immunology, 11, 614319.
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