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Sj25 c1

Manufactured by Thermo Fisher Scientific
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The SJ25-C1 is a lab equipment product manufactured by Thermo Fisher Scientific. It is a centrifuge designed for general-purpose laboratory applications. The SJ25-C1 provides reliable and efficient sample processing capabilities.

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Lab products found in correlation

Facscanto 2 flow cytometer, by BD (2 mentions) Uchl1, by Thermo Fisher Scientific (2 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Fluorescein isothiocyanate (fitc), by BD (2 mentions) Fixation buffer, by Thermo Fisher Scientific (2 mentions) Alexa fluor 647, by BioLegend (2 mentions) Brilliant violet 421, by BioLegend (2 mentions) Apc cy7, by BD (2 mentions) Pe texas red, by Thermo Fisher Scientific (2 mentions) Ber act8, by BioLegend (2 mentions) Brilliant violet 605, by BioLegend (2 mentions) Brilliant violet 650, by BioLegend (2 mentions) Percpfl710, by Thermo Fisher Scientific (2 mentions) 6 laser lsrii analytical flow cytometer, by BD (2 mentions) Alexa fluor 488, by BioLegend (2 mentions) Efluor 780, by Thermo Fisher Scientific (1 mentions) Attune nxt instrument, by Thermo Fisher Scientific (1 mentions) Lysing solution, by BD (1 mentions) Permeabilizing solution 2, by BD (1 mentions) Mhm 88, by BioLegend (1 mentions)

Close spelling variants of this product

CD19 (clone SJ25‐C1) (2 citations) CD19-Pacific blue (clone SJ25-C1) (2 citations)

7 protocols using sj25 c1

1

Nanoparticle Interaction with Blood Cells

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We investigated if the nanoparticles interact with other blood cells or components such as lymphocytes. Blood samples from healthy volunteers were exposed to anti-EpCAM nanoparticles or to PBS without nanoparticles, as described above. The samples were run over the magnetic column system. Lymphocytes were labelled by incubating the filtrates with human anti-CD19 APC-conjugated (SJ25-C1, ThermoFisher, Waltham, MA, USA; final concentration: 0.2 mg mL−1) and human anti-CD3d Alexa Fluor 48-conjugated (AB, ThermoFisher; final concentration: 0.2 mg mL−1) antibodies for 30 min at 4 °C. Red blood cells were then lysed, and samples were prepared for flow cytometry analysis, as mentioned above. Forward and side scatter area, as well as signal height, were recorded. Lymphocytes were detected using either side scatter area or forward scatter area and APC-A vs. Alexa Fluor 488-A. FACS data were processed using FlowJo V10.0.8.
Doswald S., Herzog A.F., Zeltner M., Zabel A., Pregernig A., Schläpfer M., Siebenhüner A., Stark W.J, & Beck-Schimmer B. (2022). Removal of Circulating Tumor Cells from Blood Samples of Cancer Patients Using Highly Magnetic Nanoparticles: A Translational Research Project. Pharmaceutics, 14(7), 1397.
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2

Immunophenotyping of Mesenchymal Stem Cells

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Flow cytometry was performed on an aliquot of both freshly isolated cells and on the culture-expanded MSCs between passages 1 and 5 (p1–p5), as described previously [18 , 21 ]. Freshly isolated cells were tested for CD45, CD19, CD14, and CD34 (mesenchymal fraction) using monoclonal antibodies 2D, SJ25C1, 61D3 (all ThermoFisher Scientific), and AC136 (Miltenyi Biotec), respectively. The culture-expanded MSCs were immunophenotyped using the following antibodies: anti-CD105 MEM-226 FITC (ThermoFisher Scientific), anti-CD90 DG3 FITC, and anti-CD73 AD2 APC (Miltenyi Biotec). The fixable viability dye eFluor 780 (ThermoFisher Scientific) was used to determine cell viability. Flow cytometry was performed using an Attune NxT instrument (ThermoFisher Scientific).
Čamernik K., Mihelič A., Mihalič R., Haring G., Herman S., Marolt Presen D., Janež A., Trebše R., Marc J, & Zupan J. (2020). Comprehensive analysis of skeletal muscle- and bone-derived mesenchymal stem/stromal cells in patients with osteoarthritis and femoral neck fracture. Stem Cell Research & Therapy, 11, 146.
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3

Comprehensive B Cell Immunophenotyping

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PBMCs were first stained with fluorophore-labeled antibodies against cell surface markers CD3 (OKT3, BioLegend), CD19 (SJ25-C1, Thermo Fisher), CD20 (SI Appendix, Table S2), and CD27 (O323, BioLegend), fixed (Lysing Solution, BD Biosciences), permeabilized (Permeabilizing Solution 2, BD Biosciences), and stained with fluorophore-labeled antibodies against IgG (G18-145, BD Biosciences), IgA (8E10, Miltenyi Biotec), IgD (IA6-2, BioLegend), and IgM (MHM-88, BioLegend). The stained cells were acquired on a FACS Canto II flow cytometer (BD Biosciences) and analyzed in FlowJo software v9.9.6 (BD Biosciences).
Kardava L., Rachmaninoff N., Lau W.W., Buckner C.M., Trihemasava K., Blazkova J., Lopes de Assis F., Wang W., Zhang X., Wang Y., Chiang C.I., Narpala S., McCormack G.E., Liu C., Seamon C.A., Sneller M.C., O’Connell S., Li Y., McDermott A.B., Chun T.W., Fauci A.S., Tsang J.S, & Moir S. (2022). Early human B cell signatures of the primary antibody response to mRNA vaccination. Proceedings of the National Academy of Sciences of the United States of America, 119(28), e2204607119.
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4

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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5

Flow Cytometric Analysis of Lymphocytes

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In both the human and murine experiments, the washed lymphocyte enriched pellets were suspended in 100 microliters HBSS containing a fluorescent-tagged anti-CD19 antibody. Either phycoerythrin (PE) (human experiments: Abcam ab1168 mouse anti-human; Mouse experiments: Abcam ab90753 rat anti-mouse) or APC-tagged anti-CD19 antibodies was used (Invitrogen, SJ25-C1), concentrations per manufacturers recommendations. Following a 10-minute incubation, 400 microliters of 1 micromolar Fluo-4 AM in 1xHBSS was added. The lymphocytes were allowed to load with Fluo-4 AM for 30 minutes and then the pellets were washed twice with 1x HBSS to remove excess Fluo-4 AM and fluorescent antibody. Baseline fluorescence was analyzed using a Becton Dickinson LSR II flow cytometer. Side scatter and forward scatter light were used for gating the lymphocyte population. Crossover of phycoerythrin and Fluo-4 fluorescence was compensated using single fluorophore controls. Fluorescent measurements were acquired in the absence and presence of RyR agonists, either caffeine (50mM) or 4-CmC (0.5mM).
Kushnir A., Santulli G., Reiken S.R., Coromilas E., Godfrey S.J., Brunjes D.L., Colombo P.C., Yuzefpolskaya M., Sokol S.I., Kitsis R.N, & Marks A.R. (2018). Ryanodine Receptor Calcium Leak in Circulating B-lymphocytes as a Biomarker in Heart Failure. Circulation, 138(11), 1144-1154.
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6

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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7

Flow Cytometry-Based NK Cell Cytotoxicity

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CD19 expression on the surface of target cells was analyzed by flow cytometry using FITCcoupled CD19-specific antibody (SJ25-C1; Invitrogen). Cytotoxicity of NK-92 cells towards established cancer cell lines and primary human pre-B-ALL blasts was analyzed in FACSbased assays as described [34] . Target cells were labeled with calcein violet AM (CV) (Molecular Probes, Invitrogen) and incubated with effector cells at various effector to target (E/T) ratios for 2 h at 37°C. Then 150 L of a 1 g/mL propidium iodide (PI) solution were added to each sample before flow cytometric analysis in a FACSCanto II flow cytometer (BD Biosciences). Dead target cells were identified as CV and PI double positive. Spontaneous target cell lysis in the absence of NK cells was subtracted to calculate specific cytotoxicity. Data were analyzed using FACSDiva software (BD Biosciences).
Oelsner S., Friede M.E., Zhang C., Wagner J., Badura S., Bader P., Ullrich E., Ottmann O.G., Klingemann H., Tonn T, & Wels W.S. (2017). Continuously expanding CAR NK-92 cells display selective cytotoxicity against B-cell leukemia and lymphoma. Cytotherapy, 19(2).
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