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Spherobeads

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Spherobeads are uniform, spherical beads designed for a variety of laboratory applications. They are available in a range of sizes and materials to suit different needs. The core function of Spherobeads is to provide a consistent, controlled platform for various experimental and analytical processes.

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Lab products found in correlation

Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions) Anti human hla dr allophycocyanin apc, by BD (2 mentions) Anti human hla dq fitc, by BD (2 mentions) Onecomp beads, by Thermo Fisher Scientific (2 mentions) Live dead stain, by Thermo Fisher Scientific (2 mentions) Ly6g 1a8, by BD (1 mentions) Fc block anti cd16 32, by BD (1 mentions) Cd11b m1 70, by BioLegend (1 mentions) Cd4 rm4 5, by BioLegend (1 mentions) Cd8 53 6, by BioLegend (1 mentions) Dnase 1, by Merck Group (1 mentions) Collagenase type 4, by Worthington (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions)

4 protocols using spherobeads

1

Flow Cytometry Staining Protocol

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To stain for flow cytometry, 5×105 cells per well were incubated in 1× FACS buffer (1% bovine serum albumin, 0.1% sodium azide, and 1 mM EDTA) for 20 min at 4 °C with anti-human HLA-DR:allophycocyanin/APC (1:20; BD Biosciences, San Jose, CA, USA, 559866), anti-human HLA-DQ FITC (1:20; BD Biosciences, 347453), anti-human CD14:phycoerythrin (1:100, Biolegend, no. 301806) and anti-human CD19:peridinin chlorophyll (1:100; Biolegend, no. 302228). Cells were fixed with 1% paraformaldehyde (Electron Microscopy Services, Hatfield, PA, USA) for 30 min at 4 °C. Cells were stored in 1× FACS buffer at 4 °C until run on FACS Calibur within 1 week of staining. Spherobeads (BD Biosciences) and OneComp Beads (eBiosciences) were used to set voltages and compensation settings between runs. Analysis of flow cytometry data was performed on FlowJo Software v10.0.6 (Ashland, OR, USA).
Kannarkat G.T., Cook D.A., Lee J.K., Chang J., Chung J., Sandy E., Paul K.C., Ritz B., Bronstein J., Factor S.A., Boss J.M, & Tansey M.G. (2015). Common genetic variant association with altered HLA expression, synergy with pyrethroid exposure, and risk for Parkinson’s disease: an observational and case–control study. NPJ Parkinson's Disease, 1, 15002-.
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2

Flow Cytometry of Immune Cell Subsets

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Cells were resuspended in staining buffer (PBS with 2% FCS; 5 mM EDTA), blocked with anti-CD16/32 Fc block (BD, Biosciences, Franklin Lakes, NJ, USA), and labeled with fluorochrome-conjugated antibodies for 1 h. Antibodies against CD45 (HI30; BD, Biosciences), CD3 (145-2C11; Invitrogen), CD4 (RM4-5; BioLegend), CD8 (53-6.7; BioLegend), Ly6C (HK1.4; BioLegend), Ly6G (1A8; BD, Bioscience), CD11b (M1/70; BioLegend) and NK1.1 (PK136; BD, Bioscience) were used, and a near infrared (NIR) LIVE/DEAD stain (Life Technology) was used to exclude dead cells. Counting beads (Spherobeads, BD) were added to samples before acquisition. Samples were acquired on a BD LSRFortessa flow cytometer, and data analysis was performed using FlowJo 10.7.
Ng W.H., Liu X., Ling Z.L., Santos C.N., Magalhães L.S., Kueh A.J., Herold M.J., Taylor A., Freitas J.R., Koit S., Wang S., Lloyd A.R., Teixeira M.M., Merits A., Almeida R.P., King N.J, & Mahalingam S. (2023). FHL1 promotes chikungunya and o’nyong-nyong virus infection and pathogenesis with implications for alphavirus vaccine design. Nature Communications, 14, 6605.
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3

Flow Cytometry Staining Protocol

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To stain for flow cytometry, 5×105 cells per well were incubated in 1x FACS buffer (1% bovine serum albumin, 0.1% sodium azide, 1 mM EDTA) for 20 minutes at 4°C with anti-human HLA-DR:allophycocyanin/APC (1:20, BD Biosciences 559866), anti-human HLA-DQ FITC (1:20, BD Biosciences 347453), anti-human CD14:phycoerythrin/PE (1:100, Biolegend #301806) and anti-human CD19:peridinin chlorophyll/PerCP (1:100, Biolegend #302228). Cells were fixed with 1% paraformaldehyde (Electron Microscopy Services) for 30 minutes at 4°C. Cells were stored in 1x FACS Buffer at 4°C until run on FACS Calibur within 1 week of staining. Spherobeads (BD Biosciences) and OneComp Beads (ebiosciences) were used to set voltages and compensation settings between runs. Analysis of flow cytometry data was performed on FlowJo Software v10.0.6.
Kannarkat G.T., Cook D.A., Lee J.K., Chang J., Chung J., Sandy E., Paul K.C., Ritz B., Bronstein J., Factor S.A., Boss J.M, & Tansey M.G. (2015). Common genetic variant association with altered HLA expression, synergy with pyrethroid exposure, and risk for Parkinson’s disease: an observational and case–control study. NPJ Parkinson's Disease, 1, 15002-.
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4

Lung Immune Cell Profiling by FACS

Cited 1 time
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The infected mice were culled and perfused with PBS. Lung tissues were collected for flow cytometry (fluorescence-activated cell sorting [FACS]) analysis (70 (link)). Lungs were minced and digested in type IV collagenase (0.2 mg/mL; Worthington) with DNase I (0.05 mg/mL; Sigma) and passed through 70-μm cell strainers. Cells were stained with antibodies for 45 min in FACS buffer (PBS with 0.5% [wt/vol] bovine serum albumin [BSA] and 2 mM EDTA). Fluorochrome-conjugated monoclonal antibodies against mouse CD45 (30-F11), CD3 (17A2), CD8 (53.6-7), CD4 (RM.4-5), Ly6C (HK1.4), CD11b (M1/70), Ly6G (1A8), B220 (RA3-6B2), CD11c (N418), SiglecF (E50-2440), NKp46 (29A1.4), major histocompatibility complex class II (MHC-II) (M5/114.15.2), CD64 (454-5/7.1), CCR2 (475301), and CD106 [429(MVCAM.A)] (all purchased from eBiosciences) were used, and near-infrared (NIR) LIVE/DEAD stain (Thermo Fisher) was used to exclude dead cells. Counting beads (Spherobeads; BD) were added to the samples before acquisition. Cell populations were analyzed on a BD LSRFortessa cell analyzer with BD FACSDiva software, version 6.1.3. Data analysis was performed with FlowJo (TreeStar, Inc.) software, version 9.0.
Liu X., Mostafavi H., Ng W.H., Freitas J.R., King N.J., Zaid A., Taylor A, & Mahalingam S. (2022). The Delta SARS-CoV-2 Variant of Concern Induces Distinct Pathogenic Patterns of Respiratory Disease in K18-hACE2 Transgenic Mice Compared to the Ancestral Strain from Wuhan. mBio, 13(3), e00683-22.
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