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J3 119

Manufactured by Beckman Coulter
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Sourced in France

The J3-119 is a compact benchtop centrifuge designed for general-purpose laboratory applications. It offers a maximum speed of 6,000 rpm and can accommodate a variety of sample tubes and microplates. The centrifuge is easy to operate and provides consistent, reliable performance.

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Lab products found in correlation

Navios 10 color flow cytometer, by Beckman Coulter (1 mentions) Kaluza software version 1, by Beckman Coulter (1 mentions) Immu 357, by Beckman Coulter (1 mentions) Summit 4, by Beckman Coulter (1 mentions) Cyan adp flow cytometer, by Beckman Coulter (1 mentions) Facsaria 3 cell sorter, by BD (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Gentamicin, by Thermo Fisher Scientific (1 mentions) Amphotericin b, by Thermo Fisher Scientific (1 mentions) Lsrfortessa x50, by BD (1 mentions) Rpa t8, by BioLegend (1 mentions) Hcd56, by BioLegend (1 mentions) Universal nuclease, by Thermo Fisher Scientific (1 mentions) Igg g18 145, by BD (1 mentions) Igm g20 127, by BD (1 mentions) Facs canto 2 instrument, by BD (1 mentions) Fixable viability dye, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Lonza (1 mentions) Anti epha2, by R&D Systems (1 mentions)

Spelling variants of this product

CD19-ECD (J3-119) (7 citations) CD19 (J3-119, (4 citations) PE-Cy5 anti-CD19 (J3-119) (3 citations) CD19-PC7 (clone J3-119, (3 citations) CD19-PE-Cy7 (clone J3-119; (3 citations) Anti-human CD19-Pc7 (clone J3-119; (2 citations) Anti-CD19 (J3-119) (2 citations)

5 protocols using j3 119

1

Whole Blood Immunophenotyping in FSHD

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As a second step, we measured the whole blood immunophenotyping of a subset of 15 FSHD patients and 15 new sex-age healthy controls. Samples were not pre-selected out of an initial pool. Peripheral fresh whole blood samples were analysed by flow cytometry as described by Aguirre-Gamboa et al. [28 (link)]. Briefly, samples were stained with a antibody cocktail containing the following fluorochrome conjugated antibodies (dilution, cone, distributor): CD45-KO (1:50, J33, Beckman Coulter), CD16-FITC (1:50, 3G8, Beckman Coulter), HLA-DR-PE (1:10, immu-357, Beckman Coulter), CD14-ECD (1:100, UCHT1, Beckman Coulter), CD4-PE-Cy5.5 (1:200, 13B8.2, Beckman Coulter), CD25-PC7 (1:50, M-A251, Becton & Dickinson), CD56-APC (1:50, N901, Beckman Coulter), CD8-APC-AF700 (1:400, B9.11, Beckman Coulter), CD19-APC-AF750 (1:50, J3-119, Beckman Coulter), CD3-PB (1:50, UCHT1, Beckman Coulter). Surface expression was measured on a 10-color Navios flow cytometer (Beckman Coulter) and analysed using Kaluza software version 1.3 (Beckman Coulter) (see Supplemental Figure 2 for gating strategy).
Greco A., Mul K., Jaeger M.H., dos Santos J.C., Koenen H., de Jong L., Mann R., Fütterer J., Netea M.G., Pruijn G.J., van Engelen B.G, & Joosten L.A. (2024). IL-6 and TNF are Potential Inflammatory Biomarkers in Facioscapulohumeral Muscular Dystrophy. Journal of Neuromuscular Diseases, 11(2), 327-347.
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2

Monocyte Phenotypic Validation by Flow

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Phenotypic validation of monocytes isolated from healthy donors was performed by flow cytometry analysis using the PE‐conjugated anti‐CD14 mouse monoclonal antibody TÜK4 (Miltenyi Biotec, SAS, Paris, France), the FITC‐conjugated anti‐CD3 mouse monoclonal antibody UCHT1 (Becton Dickinson France, SAS, Le Pont de Claix, France) and the PE‐conjugated anti‐CD19 mouse monoclonal antibody J3.119 (Beckman Coulter France, SAS, Villepinte, France). All FACS analyses were performed using a Cyan ADP flow cytometer equipped with Summit 4.3 software (Beckman Coulter France, SAS, Villepinte, France).
Galiègue‐Zouitina S., Fu Q., Carton‐Latreche C., Poret N., Cheok M., Leprêtre F., Figeac M., Quesnel B., El Bouazzati H, & Shelley C.S. (2021). Bimodal expression of RHOH during myelomonocytic differentiation: Implications for the expansion of AML differentiation therapy. EJHaem, 2(2), 196-210.
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3

Cell Sorting and Functional Assay Protocol

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For functional studies, cells were sorted on a BD FACS Aria III cell sorter (BD Biosciences). Cells were stained for anti-CD19-APC (J3-119, Beckman Coulter; EphA2-CAR) or anti-mouse IgG F(ab')2 fragment specific-AlexaFluor 647 (HER2-CAR) plus anti-CD116-PE (4H1, BioLegend or hGM-CSFR-M1, BD Biosciences). DAPI (Thermo Fisher) was used as a viability indicator. Cells were rested 48-72 hours in RPMI containing 20% FBS, 25 μg/ml gentamicin (Gibco), 1X penicillin-streptomycin (Gibco), and 1.5 μg/ml amphotericin B (Gibco) prior to functional assays.
Lange S., Sand L.G., Bell M., Patil S.L., Langfitt D, & Gottschalk S. (2021). A chimeric GM-CSF/IL18 receptor to sustain CAR T-cell function. Cancer discovery, 11(7), 1661-1671.
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4

Assessing HA-Directed B Cell Binding

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In order to assess HA- directed B cell binding and cross-reactivity, peripheral blood mononuclear cells (PBMC’s) from samples with different stem binding based on ELISA assay (n = 18) were further analyzed by flow cytometry. Three groups were selected for the analysis: negative for stem antibodies (n = 5), reactive to one stem only (n = 4), and samples with cross-reactive stem binding (n = 9). Cryopreserved PBMC’s were thawed in R-10 media containing 50 U/mL of Universal Nuclease (ThermoFisher). Cells were washed and resuspended in PBS for staining with UV-Blue viability dye (ThermoFisher) for 20 minutes at room temperature. After washing, surface staining was performed using antibodies against IgM (G20-127, BD Biosciences), IgG (G18-145, BD Biosciences), CD8 (RPA-T8, BioLegend), CD3 (OKT3, BioLegend), CD56 (HCD56, BioLegend), CD14 (M5E2, BioLegend), CD19 (J3-119, Beckman Coulter), CD27 (O323, BioLegend), CD38 (HIT2, BioLegend), and HA probes27 –29 (link). H1 NC 99 and H5 INDO 05 HAs were expressed and biotinylated followed by fluorochrome labeling as previously described29 (link). Stained cells were run on a BD LSRFortessa X-50 and data analysis was performed using FlowJo (TreeStar). The gating strategy is demonstrated in supplemental Fig. 2. Statistical significance at 95% confidence interval was done using two-tailed t-test.
Yassine H.M., McTamney P.M., Boyington J.C., Ruckwardt T.J., Crank M.C., Smatti M.K., Ledgerwood J.E, & Graham B.S. (2018). Use of Hemagglutinin Stem Probes Demonstrate Prevalence of Broadly Reactive Group 1 Influenza Antibodies in Human Sera. Scientific Reports, 8, 8628.
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5

Comprehensive Flow Cytometry Analysis of CAR T-cells

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The FACSCanto II instrument (BD Biosciences) was used for acquisition and FlowJo v10 (FlowJo) was used for analysis of flow cytometry data. Samples were washed with and stained in PBS (Lonza) with 1% FBS. For all experiments, known negatives (e.g. NT T-cells) served as gating controls. eBioscience Fixable viability dyes (Invitrogen) were used to exclude dead cells from analysis. T-cells were evaluated for CAR expression 5-10 days post transduction using anti-CD19 (J3-119, Beckman Coulter; SJ25C1, BD Biosciences) or anti-human IgG F(ab')2 fragment specific antibody (Jackson ImmunoResearch) for EphA2-CAR. Anti-mouse IgG F(ab')2 fragment specific antibody (Jackson ImmunoResearch) was used to detect HER2-CAR expression. GM18 expression was analyzed by staining with anti-CD116 (4H1, BioLegend). CAR T-cell phenotype was established using the following antibodies: CD4 (SK3, BD Biosciences), CD8 (HIT8a, BD Biosciences), CD45RA (HI100, BD Biosciences), and CCR7 (G043H7, BioLegend; 150503, BD Biosciences). The beta chain of GM18 was detected with anti-CD131 (3D7, BD Biosciences). Cell surface EphA2 expression was detected using anti-EphA2 (371805, R&D Systems).
Lange S., Sand L.G., Bell M., Patil S.L., Langfitt D, & Gottschalk S. (2021). A chimeric GM-CSF/IL18 receptor to sustain CAR T-cell function. Cancer discovery, 11(7), 1661-1671.
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