Summit data acquisition and analysis software

Manufactured by Agilent Technologies

The Summit Data Acquisition and Analysis Software is a comprehensive data management solution developed by Agilent Technologies. The software is designed to provide users with the tools to efficiently acquire, analyze, and manage data generated from various laboratory instruments and experiments.

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Lab products found in correlation

Lsrfortessa flow cytometer, by BD (2 mentions) Tnf α pe, by BD (2 mentions) Streptavidin pacific blue, by Thermo Fisher Scientific (2 mentions) Ifn γ apc 4s b3, by BD (2 mentions) Cd49d 9f10, by BD (2 mentions) Cd4 bv510 l200, by BD (2 mentions) Ifn γ brilliant violet 711 4s b3, by BioLegend (2 mentions) Perforin biotinylated pf 344, by Mabtech (2 mentions) Tnf α pe cy7, by BD (2 mentions) Pacific blue conjugated anti cd3 clone sp34 2, by BD (2 mentions) Cyan adp flow cytometer, by Agilent Technologies (2 mentions) Cd3 pb sp34 2, by BD (1 mentions) Cd8 pb rpa t8, by BD (1 mentions) Ebio64cap17, by Thermo Fisher Scientific (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Cd8 pacificblue rpa t8, by BD (1 mentions) Cd3 pacific blue sp34 2, by BD (1 mentions)

Close spelling variants of this product

Data Analysis software (2 citations) Summit software (38 citations)

6 protocols using summit data acquisition and analysis software

Assessing T-cell Activation and IFN-γ Production in TB Patients

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PBMCs isolated from TB patients were cultured in at 37°C in a humidified atmosphere with 5% CO2, and stimulated with Lithium chloride (LiCl, Sigma) at 20mM for 24 hours. Then, cells were collected and stained with Pacific blue-conjugated anti-human CD3 (Clone SP34–2, BD), FITC-conjugated anti-human CD14 (Clone HCD14, Bioelgend), BV605-conjugated anti-human CD69 (clone FN50, Biolegend), BV711-conjugated anti-human IFN—γ (Clone 4S.B3, Biolegend), APC-conjugated anti-human betta-catenin (Clone 196624, R&D). After staining, cells were fixed with 2% formaldehyde-PBS (Protocol Formalin, Kalamazoo, MI) and subjected to run on a BD LSRF ortessa flow cytometer (BD Biosciences, Qume Drive, San Jose, CA). Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (Dako Cytomation).

Fan L., Shen H., Huang H., Yang R, & Yao L. (2017). Impairment of Wnt/β-catenin signaling in blood cells of patients with severe cavitary pulmonary tuberculosis. PLoS ONE, 12(3), e0172549.

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Flow Cytometry Antibody Staining Protocol

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The following antibodies were used for culturing or surface and intracellular cytokine staining (ICS) for flow cytometry: CD28 (CD28.2, BD), CD49d (9F10, BD), CD3-PB (SP34-2, BD), CD4-BV510 (L200, BD), CD8-PB (RPA-T8, BD), IFN-γ-APC (4S.B3, BD), IFN-γ Brilliant Violet 711 (4S.B3, Biolegend), TNF-α-PE (MAb11, BD), TNF-α-PE-Cy7 (MAb11, BD), IL-17-PE (eBio64CAP17, eBioscience), IL-22-biotinylated (anti-human IL-22, RD), Streptavidin-Pacific blue (Invitrogen), Perforin-biotinylated (Pf-344, Mabtech), Caspase 3-AF647 (C92-605,BD), and anti-Vγ2-FITC (7A5, Pierce).
After staining, the cells were fixed and subjected to flow cytometry. Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (DakoCytomation).

Yang E., Yang R., Guo M., Huang D., Wang W., Zhang Z., Chen C., Wang F., Ho W., Shen L., Xiao H., Chen Z.W, & Shen H. (2018). Multidrug-resistant tuberculosis (MDR-TB) strain infection in macaques results in high bacilli burdens in airways, driving broad innate/adaptive immune responses. Emerging Microbes & Infections, 7, 207.

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Flow Cytometry Analysis of T-cell Subsets

Cited 2 times

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This was done as we previously described [24 (link), 33 (link), 44 (link)]. Cells were harvested at day 7 after culture and stained with Pacific blue-conjugated anti-CD3 (Clone SP34-2, BD, Franklin Lakes, NJ), PE conjugated anti-CD4 (clone OKT4, eBioscience, San Diego, CA), FITC-conjugated Vγ2 (Clone 7A5, Pierce, Rockford, IL) and purified Vδ2 (clone 15D, Pierce) in combination of allophycocyanin-conjugated goat anti mouse IgG (Dako, Carpinteria, CA). After staining, cells were fixed with 2% formaldehyde-PBS (Protocol Formalin, Kalamazoo, MI) and subjected to run on a CyAn ADP flow cytometer (DakoCytomation, Carpinteria, CA). Lymphocytes were gated based on forward- and side-scatters, and pulse width and at least 40 000 gated events were analyzed by using Summit Data Acquisition and Analysis Software (Dako Cytomation). Further special gates and quadrants for data analysis were determined on nonstaining, specific antibody staining, and isotype control antibody background staining.

Shen H., Wang Y., Chen C.Y., Frencher J., Huang D., Yang E., Ryan-Payseur B, & Chen Z.W. (2015). Th17-related cytokines contribute to recall-like expansion/effector function of HMBPP-specific Vγ2Vδ2 T cells after Mycobacterium tuberculosis infection or vaccination. European journal of immunology, 45(2), 442-451.

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Multiparametric Flow Cytometry Analysis

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This was done according to our previously described [34 (link)]. Cells were stained with Pacific blue-conjugated anti-CD3 (Clone SP34–2, BD, Franklin Lakes, NJ), FITC-conjugated anti-CD19(clone HIB19, Biolegend, San Diego, CA), APC-conjugated anti-CD27(clone O323, Biolegend, San Diego, CA), BV605-conjugated anti-CD69(clone FN50, Bio-legend, San Diego, CA), AF700-conjugated anti-CD138(clone MI15, Biolegend, San Diego, CA), PerCP/Cy5.5-conjugated anti-CXCR5 (CloneJ252D4, Biolegend, San Diego, CA), BV510-conjugated anti-CD4 (Clone A161A1, Biolegend, San Diego, CA). After staining, cells were fixed with 2% formaldehyde-PBS (Protocol Formalin, Kalamazoo, MI) and subjected to run on a BD LSRFortessa flow cytometer (BD Biosciences, Qume Drive, San Jose, CA). Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (Dako Cytomation).

Huang H., Qian X., Pan R., Shen L., Liang S., Wang F., Zhang P., Shen H, & Chen Z.W. (2018). 23-valent pneumococcal polysaccharide vaccine elicits hierarchical antibody and cellular responses in healthy and tuberculosis-cured elderly, and HIV-1-infected subjects. Clinical immunology (Orlando, Fla.), 193, 1-9.

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Flow Cytometric Analysis of Lymphocyte Activation

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FITC-TCR Vδ2+ antibodies (BD Bioscience) were used to analyze 100 μl samples of anticoagulated blood from the 32 patients. After 10 μl of Phycoerythrin (PE)- FasL, PE-CY7- Fas, CD3-PB antibodies (BD Bioscience) was added, the whole blood samples were then incubated at room temperature for 30 min in the dark. After 1 ml red blood cell lysis buffer was added, cells were incubated at room temperature in the dark for an additional 15–20 min. After vortexing, the supernatant was discarded and the suspensions were centrifuged at 1400 rpm for 5 min. After using PBS (Protocol Formalin, Kalamazoo, MI) to wash again, the remaining cells were then resuspended in 400 μl PBS. Lymphocyte populations were gated based on the forward and side-scatters on a CyAn ADP flow cytometer (Dako Cytomation, Carpinteria, CA). Gated events were analyzed using Summit Data Acquisition and Analysis Software (Dako Cytomation)(Additional files 1, 2, 3 and 4).

Yan L., Shen H, & Xiao H. (2018). Characteristics of peripheral Vγ2Vδ2 T cells in interferon-γ release assay negative pulmonary tuberculosis patients. BMC Infectious Diseases, 18, 453.

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Cytokine Analysis by Flow Cytometry

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The following Abs were used for culture or surface and intracellular cytokine staining for flow cytometry: CD28 (CD28.2, BD), CD49d (9F10, BD), CD3-Pacific blue (SP34-2,BD), CD4-BV510 (L200, BD), CD8- Pacific blue (RPA-T8, BD), IFN-γ-APC (4S.B3, BD), IFN-γ Brilliant Violet 711 (4S.B3, Biolegend), TNF-α-PE (Mab11, BD), TNF-α-PE-Cy7 (Mab11, BD), IL-17-PE (eBio64CAP17, eBioscience), IL-22-biotinylated (anti-human IL-22, RD), Streptavidin-Pacific blue (invitrogen), Perforin-biotinylated (Pf-344, Mabtech), Caspase 3-AF647 (C92-605,BD), anti-Vγ2-FITC (7A5, Pierce).
After staining, cells were fixed and subjected to analysis on flow cytometer of BD LSRFortessa^TM Cell Analyzer. Lymphocytes were gated based on forward- and side-scatters, and at least 40,000 gated events were analyzed using Summit Data Acquisition and Analysis Software (Dako Cytomation).

Shen H., Yang E., Guo M., Yang R., Huang G., Peng Y., Sha W., Wang F, & Shen L. (2022). Adjunctive Zoledronate + IL-2 administrations enhance anti-tuberculosis Vγ2Vδ2 T-effector populations, and improve treatment outcome of multidrug-resistant tuberculosis1. Emerging Microbes & Infections, 11(1), 1790-1805.

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