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Ccr7 bv510

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CCR7-BV510 is a fluorescent-labeled antibody that binds to the CCR7 receptor. CCR7 is a chemokine receptor that plays a role in the trafficking and homing of T cells and dendritic cells. The BV510 fluorophore allows for the detection and analysis of CCR7-expressing cells using flow cytometry.

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Lab products found in correlation

Cd3 fitc, by BD (2 mentions) Lsrfortessa, by BD (2 mentions) Cd3 apc cy7, by BD (1 mentions) Ficoll, by Merck Group (1 mentions) Histopaque, by Merck Group (1 mentions) Cd4 apc h7, by BD (1 mentions) Cd4 bv421, by BD (1 mentions) Cd45ra apc, by BD (1 mentions) Cd8 bv605, by BD (1 mentions) Cd16 apc, by BD (1 mentions) Cd3 pe cy7, by BD (1 mentions) Cd56 fitc, by BD (1 mentions) Cd45 buv395, by BD (1 mentions) Zombie nir, by BioLegend (1 mentions) Nkp30 bv711, by BD (1 mentions) Cxcr3 bv421, by BD (1 mentions) Nkp44 percp efluor 710, by Thermo Fisher Scientific (1 mentions) Cxcr1 pe, by R&D Systems (1 mentions) Cd3 pe vio770, by Miltenyi Biotec (1 mentions) Cd56 bv510, by BD (1 mentions)

4 protocols using ccr7 bv510

1

Immunophenotyping of Cryopreserved PBMCs

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Peripheral blood mononuclear cells (PBMC) were obtained from whole blood using Histopaque® and Ficoll (Sigma-Aldrich, Saint Louis, MO, USA) different density gradients. These cells were cryopreserved, and then thawed at the time of each assay. Then, was used a concentration of 2 × 105 viable cells/mL, and submitted to immunophenotyping assay with surface antibodies for 20 min at 2–8 °C. After, the cells were washed with phosphate buffer plus fetal bovine serum (FBS) (PBS pH 7.4 at 2% FBS), and centrifuged at 400× g for 5 min. After centrifugation, cells were fixed with 1% paraformaldehyde solution and subsequently acquired in a flow cytometer (LSR FortessaTM, BD Biosciences, Franklin Lakes, NJ, USA). The analysis was performed using Flow Jo software v10.6 (BD Biosciences).
The anti-human antibodies used in the immunophenotyping assay were: panel I (activation)-CD3-FITC, CD4-APCH7, CD8-BV605, CD38-PECy7, OX40-BV711 and panel II (memory)–CD3-APC-Cy7, CD4-BV421, CD8-BV605, CD45RA-APC, CCR7-BV510 (BD Biosciences).
Pacheco V., Cuber Guimarães R., Corrêa-Moreira D., Magalhães C.E., Figueiredo D., Guttmann P., Trindade G.F., da Silva J.F., Ano Bom A.P., de Lourdes Maia M., Melgaço J.G., da Costa Barros T.A., da Silva A.M, & Oliveira M.M. (2023). Clinical Profile of SARS-CoV-2 Infection: Mechanisms of the Cellular Immune Response and Immunogenetic Markers in Patients from Brazil. Viruses, 15(7), 1609.
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2

Multiparametric Characterization of NK Cells

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One-step staining of cell-surface antigens was performed using fluorochrome-conjugated primary antibodies as previously described (10 (link), 18 (link), 33 (link)). For the analysis of blood NK cells two antibody panels were constructed around CD56-FITC, CD16-APC, and CD3-PE-Cy7 (BD Bioscience, San Jose, CA) antibodies. NK cell activation receptors were evaluated with CD69-BV421, NKp30-BV711 (BD Bioscience), and NKp44-PerCP eFluor 710 (eBioscience, San Diego, CA) antibodies. Chemokine expression levels were tested using CXCR1-PE (R&D Systems, Minneapolis, MN), CXCR3-BV421, and CCR7-BV510 (BD Bioscience) antibodies. For the TINK analysis, Zombie NIR (BioLegend), CD45 BUV395, CD56 BV510, CD16 BUV737 (BD Bioscience), and CD3 PE-Vio770 (Miltenyi Biotec) antibodies were used. Suitable IgG controls were acquired from the same vendors. FACS analyses were performed using the BD LSRFortessa™ cell analyzer, and analyzed using FlowJo v10 (FlowJo, LLC; Ashland, OR) software.
Vujanovic L., Chuckran C., Lin Y., Ding F., Sander C.A., Santos P.M., Lohr J., Mashadi-Hossein A., Warren S., White A., Huang A., Kirkwood J.M, & Butterfield L.H. (2019). CD56dim CD16− Natural Killer Cell Profiling in Melanoma Patients Receiving a Cancer Vaccine and Interferon-α. Frontiers in Immunology, 10, 14.
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3

Multiparametric Flow Cytometry of T-Cell Subsets

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Cells were first stained with CD3-FITC, CD4-PE, CD8-APC-Cy7, CD45RA PE-Cy7, CD45RO-APC, CCR7-BV510, HLA-DR-BV605, and TCRγδ-BV421 (BD Biosciences). For intracellular staining with IFNγ-PE-Cy7, TNF-AL700, IL-2-BV421, and IL-17-BV510, cells were fixed and permeabilized using the BD Biosciences Cytofix/Cytoperm Kit. Data were acquired on BD LSRFortessa® (50,000 events), and cell frequencies, as well as median fluorescence intensity (MFI), were measured using FlowJo 10 software (Tree Star Inc.). Supernatants of PBMC cultures were collected and stored at -20°C for cytokine assays. Concentrations of IFNγ and IL-10 were measured by ELISA (R&D Systems) according to the manufacturer’s instructions.
Sarno A., Leite A., Augusto C., Muller I., de Ângelis L., Pimentel L., Queiroz A, & Arruda S. (2024). Impaired macrophage and memory T-cell responses to Bacillus Calmette-Guerin nonpolar lipid extract. Frontiers in Immunology, 14, 1263352.
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4

Immunophenotyping of Severe COVID-19 PBMC

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Cryopreserved PBMC from patients hospitalized because of COVID-19 severe interstitial pneumonia (severe) and OTD subjects were thawed, washed and rested for 30 min in complete RPMI-1640 medium supplemented with 10% FCS, 2mM L-glutamine and antibiotic antimycotic solution (100 U/ml penicillin, 0.1 μg/ml streptomycin, 0.25 μg/ml amphotericin B (Sigma- Aldrich, St. Louis, MO, USA) (Complete Medium). Subsequently, PBMC were washed and stained for flow cytometric analysis using the following fluorochrome conjugated antibodies: CD8 BV421, CCR7 BV510, PD1 PE-CF594, CD45RA BV650, CD56 BV786, TIM-3 BB515, CD69 PE, CD4 BB700, CD3 APC-H7 (BD Biosciences, San Diego, CA, USA). After fixation and permeabilization (Fixation/Permeabilization Solution Kit; BD Biosciences), cells were stained with anti-Granzyme B Alexa 647 (BD Biosciences). To detect circulating T follicular helper (TFH) cells 1×106 PBMC were stained with the following antibodies: CD3 BV510, CD4 BB700, CD8 BV786, PD1 PE and CXCR5 BV421. The following antibodies were used to identify plasma cells: CD19 BV605, CD27 BB515, CD38 BV421and CD138 BV480. Flow cytometry was performed with a 12-color Celesta (BD Biosciences) instrument and data analyzed with FlowJo 10 software.
Mele D., Calastri A., Maiorano E., Cerino A., Sachs M., Oliviero B., Mantovani S., Baldanti F., Bruno R., Benazzo M., Grifoni A., Sette A., Mondelli M.U, & Varchetta S. (2021). High Frequencies of Functional Virus-Specific CD4+ T Cells in SARS-CoV-2 Subjects With Olfactory and Taste Disorders. Frontiers in Immunology, 12, 748881.
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