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Accucheck

Manufactured by Thermo Fisher Scientific
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The Accucheck is a laboratory instrument designed for precise measurement and analysis. It provides accurate and reliable quantitative results for various applications in the scientific and research fields.

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Lab products found in correlation

Cd54 fitc, by Bio-Rad (1 mentions) Cd106 fitc, by Bio-Rad (1 mentions) Cd142 fitc, by Bio-Rad (1 mentions) Cd31 fitc, by Bio-Rad (1 mentions) Cd105 fitc, by Bio-Rad (1 mentions) Facscalibur, by BD (1 mentions) Anti ly 6g rb6 8c5, by BioLegend (1 mentions) Ovalbumin (ova), by Worthington (1 mentions) Nod scid il2rgammanull nsg mice, by Jackson ImmunoResearch (1 mentions) X rad 320, by Precision X-Ray (1 mentions) Anti cy5 af647 microbeads, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

AccuCheck Counting Beads AccuCheck beads AccuCheck microbeads AccuCheck fluorescent beads Fluorescent AccuCheck counting beads

7 protocols using accucheck

1

Flow Cytometry Analysis of Cell-Derived Particles

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Flow cytometry for assessment of cell surface and MP expression from cancer cells lines and HUVEC was performed on a FACSCalibur (Becton Dickinson, Oxford, UK). For assessment of cell free media was obtained by centrifugation at 400g for 5 minutes to pellet cells. 25µl of this media was then incubated with 5µl of antibody (CD31:FITC, CD54:FITC, CD105:FITC, CD106:FITC or CD142:FITC) (Bio-Rad, Hemel Hempstead, UK) at room temperature in the dark for 30 minutes, when counting beads (Accucheck, Thermo-Fisher) and 150µl filtered PBS were added prior to assessment by flow cytometry using ISTH guidelines for enumeration of MP.
For assessment of cancer or HUVEC cells, 5µg of antihuman antibodies (CD31:FITC, CD54:FITC, CD105:FITC, CD106:FITC or CD142:FITC, Bio-Rad, Hemel Hempstead, UK) or an isotype matched negative control (IgG1:FITC, Bio-Rad) was then added to 50µl of cells (2x10 5 ) and incubated for 30 minutes, at room temperature in the dark. Cells were then washed in PBS and centrifuged. The pellet was then resuspended in 200µl filtered PBS and assessed by flow cytometry.
Faulkner L.G., Alqarni S., Maraveyas A, & Madden L.A. (2020). Isolated tumour microparticles induce endothelial microparticle release in vitro. Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis, 31(1).
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2

Quantifying Neutrophils in Blood and Peritoneum

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The absolute number of neutrophils were determined in freshly harvested blood and peritoneal lavage of mice. Briefly, 100 μL of blood or peritoneal lavage was stained for neutrophils and the absolute counting was performed on a flow cytometer using a bead counting method (AccuCheck, Thermo Scientific, Waltham, MA, USA) as we have previously described [12 (link),16 (link)]. Antibodies used to identify neutrophils included anti-Mac-1 (M1/70), anti-F4/80 (BM8) and anti-Ly-6G (RB6-8C5) (BioLegend, San Diego, CA, USA). The appropriate isotype-matched negative control antibodies were purchased from the same source. Bacterial counts were determined by plating 10-fold serial dilutions of blood or peritoneal lavage fluid on brain heart infusion agar plate (Becton Dickinson, Sparks, MD, USA), as previously described [16 (link)].
Mishra H.K., Ma J., Mendez D., Hullsiek R., Pore N, & Walcheck B. (2020). Blocking ADAM17 Function with a Monoclonal Antibody Improves Sepsis Survival in a Murine Model of Polymicrobial Sepsis. International Journal of Molecular Sciences, 21(18), 6688.
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3

Antigen-Specific T Cell Activation by DCs

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Primary splenic DCs were purified as previously described43 (link). Activated primary DCs were incubated for 14–16 h in the presence of 0.5 µM CpG Type B 1688 (Geneworks). Unstimulated DCs were pulsed with 100 μg OVA (Worthington) together with 5 µM CpG, and activated splenic DCs pulsed with 100 μg OVA for 45 minutes at 37 °C. Excess OVA was removed by washing. Both unstimulated and activated DCs were pulsed with BP or BP-OVA beads that remained in culture. 5 × 104 CTV-labelled purified OT-I or OT-II T cells were cultured in vitro with unstimulated or activated DCs in RPMI 1640 supplemented with 10% FBS, 100 IU/mL penicillin (MPU) and 100 μg/ml streptomycin (MPU). Cells were incubated for 60 - 64 h (37 °C, 10% CO2). T cells were stained with mAbs specific for: CD4 (GK1.5, FITC, 1:500, 100406), CD8 (53-6.7, FITC, 1:400, WEHI), TCRVα2 (B20.1, APC, 1:800, WEHI) and Ly5.1 (A20.1, PE-Cy7, 1:200, 110730) (Biolegend). T cell proliferation was analyzed using flow cytometry and CTV dilution. Counting beads (Accucheck, Thermo Fischer) were included to enable determination of the number of proliferating T cells.
Jenika D., Pounraj S., Wibowo D., Flaxl L.M., Rehm B.H, & Mintern J.D. (2024). In vivo assembly of epitope-coated biopolymer particles that induce anti-tumor responses. NPJ Vaccines, 9, 18.
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4

Xenogeneic Adoptive Transfer Model

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The xenogeneic adoptive transfer model was performed as we have previously described (10 (link)). NOD-scid IL2Rgammanull (NSG) mice (stock number is 005557 from Jackson Laboratory, Bar Harbor, ME) were housed in a specific pathogen-free facility. Weight matched (26-30g) female mice were subjected to whole-body preconditioning irradiation (225 cGy using an X RAD 320, Precision X-ray, North Branford, CT, USA) for consistent NK cell engraftment. Freshly, enriched human NK cells underwent initial overnight incubation in B0 media [DMEM, Ham’s F12 with 10% human AB serum, Pen/Strep (1%), 2-ME (20 μm), ethanolamine (50 μm), ascorbic Acid (10 μg/ml) and sodium selenite (1.6 ng/ml)] containing 2.5 ng/ml rhIL-15 (NCI), and 4x106 cells were injected via tail vein in each mouse. Mice were also administered rhIL-15 (NCI) ip at dose of 5 μg. The indicated mAbs were ip administered at a dose of 10 mg/kg. A schematic of the treatment schema is provided in Figure 2A. Blood was collected via retro-orbital route in heparin. Absolute counting of human NK cells in the peripheral blood was performed on a flow cytometer using a bead counting method (AccuCheck, Thermo Scientific, Waltham, MA, USA) according to the manufacturer’s instructions.
Mishra H.K., Dixon K.J., Pore N., Felices M., Miller J.S, & Walcheck B. (2021). Activation of ADAM17 by IL-15 Limits Human NK Cell Proliferation. Frontiers in Immunology, 12, 711621.
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5

SARS-CoV-2 S1-RBD Tetramer Assay

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PBMCs were incubated with 10 nM decoy SA-PE-AF647 for 10 minutes and then with 5 nM S1-RBD/SA-AF647 tetramer for 45 minutes at room temperature. The samples were washed and incubated with 25 μL Cy5/AF647 antibody-conjugated MicroBeads (Miltenyi, 130-091-395) for 15 minutes at 4°C, washed again, and then passed over magnetized Miltenyi LS columns. The cells that flowed through the columns were saved. The columns were then washed 3 times and removed from the magnetic field to allow elution of the bound cells in 5 mL sorter buffer. Fluorescent beads (AccuCheck, Life Technologies, PCB100) were added for the purpose of calculating the number of live lymphocytes.
Pape K.A., Dileepan T., Matchett W.E., Ellwood C., Stresemann S., Kabage A.J., Kozysa D., Evert C., Matson M., Lopez S., Krueger P.D., Graiziger C.T., Vaughn B.P., Shmidt E., Rhein J., Schacker T.W., Bold T.D., Langlois R.A., Khoruts A, & Jenkins M.K. (2022). Boosting corrects a memory B cell defect in SARS-CoV-2 mRNA–vaccinated patients with inflammatory bowel disease. JCI Insight, 7(12), e159618.
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6

Cytokine Sensitivity Assays for Mouse Hematopoietic Cells

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CFC assays and LTCs were performed as described previously (Hogge et al., 1996 (link)), using for the LTCs a mixture of irradiated M210B4 and Sl/Sl mouse stromal cells producing human SF, IL-3, FL, and G-CSF. Live (propidium iodide negative [PI−]) nonadherent cells were enumerated by flow cytometry using fluorescent counting beads (AccuCheck, Life Technologies) and their morphology determined on Wright-Giemsa-stained cytospin preparations. For suspension culture experiments, test cells were incubated in either serum-free medium (SFM) with growth factors (as for lentiviral exposure; Figures 4H–4J) or H5100 media with GM-CSF (Figure S3C).
To compare the cytokine sensitivity of IK6- and control-transduced lineage-SCA-1+CD3− B6 mouse BM cells, 104 of each were cocultured in 100 μl SFM in 96-well plates with added mouse SF or IL-3, as indicated. After 4 days, total viable (PI−) GFP+ and YFP+ cells in each well were enumerated by flow cytometry using fluorescent counting beads.
Beer P.A., Knapp D.J., Kannan N., Miller P.H., Babovic S., Bulaeva E., Aghaeepour N., Rabu G., Rostamirad S., Shih K., Wei L, & Eaves C.J. (2014). A Dominant-Negative Isoform of IKAROS Expands Primitive Normal Human Hematopoietic Cells. Stem Cell Reports, 3(5), 841-857.
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7

SARS-CoV-2 S1 RBD Tetramer Labeling

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Mouse spleen and lymph nodes or human PBMC suspensions were mixed with 1 μL of 1 μM decoy SA-PE-AF647 and incubated for 10 minutes at room temperature, and then with 0.5 μl of 1 μM wild-type S1-RBD/SA-AF647 tetramer or 1 μM wild-type S1-RBD/SA-AF647 tetramer plus 1 μM B.1.351 S1-RBD/SA-BV650 tetramer for 45 minutes at room temperature. Samples were washed with sorter buffer and 25 μl of anti-Cy5/AF647 MicroBeads (Miltenyi #130-091-395) were added and incubated for 15 minutes at 4°C. The cell suspensions were then passed over magnetized LS columns. The columns were washed 3 times to remove unlabeled cells and the cells that flowed through the columns were saved. After the last wash, the columns were removed from the magnetic field, and bound cells were eluted in 5 mL of sorter buffer. Fluorescent beads (AccuCheck, Life Technologies #PCB100) were used to calculate the number of live lymphocytes in each cell suspension.
Pape K.A., Dileepan T., Kabage A.J., Kozysa D., Batres R., Evert C., Matson M., Lopez S., Krueger P.D., Graiziger C., Vaughn B.P., Shmidt E., Rhein J., Schacker T.W., Khoruts A, & Jenkins M.K. (2021). High-affinity memory B cells induced by SARS-CoV-2 infection produce more plasmablasts and atypical memory B cells than those primed by mRNA vaccines. Cell Reports, 37(2), 109823.
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