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S6 ultra immunoscan reader

Manufactured by Cellular Technology
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The S6 ultra immunoscan reader is a laboratory instrument designed for the quantitative analysis of immunoassays. Its core function is to measure and analyze the signals generated from immunological reactions, providing accurate and reliable data for research and diagnostic purposes.

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Lab products found in correlation

Immunospot software, by Cellular Technology (4 mentions) Immunospot 5, by Cellular Technology (2 mentions) Mouse ifn γ elispot kit, by Dakewe (1 mentions) Human ifn γ elispot kit, by Dakewe (1 mentions) Sars cov 2 s peptides pool, by GenScript (1 mentions)

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ImmunoScan (3 citations)

7 protocols using s6 ultra immunoscan reader

1

IFN-γ Detection in T Cells

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IFN-γ–secreting T cells stimulated by MANA/WT peptide/scramble peptide/no peptide were detected by Human IFN-γ ELISpot assay kits (3512-4APW-2, Mabtech) according to the manufacturer’s protocol. PBMCs were plated in triplicate at 3 × 105 per well and then incubated for 24 hours. Spots were then counted using an S6 ultra immunoscan reader (Cellular Technology Ltd.), and the number of IFN-γ–positive T cells was calculated by ImmunoSpot 5.1.34 software (Cellular Technology Ltd.).
Han J., Yu R., Duan J., Li J., Zhao W., Feng G., Bai H., Wang Y., Zhang X., Wan R., Xu J., Wang X., Guan Y., Xia X., Yao Z., Fei K., Carbone D.P., Wang Z, & Wang J. (2021). Weighting tumor-specific TCR repertoires as a classifier to stratify the immunotherapy delivery in non–small cell lung cancers. Science Advances, 7(21), eabd6971.
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2

ELISpot Assay for CAR-T Cell Function

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For enzyme-linked immunosorbent spot (ELISpot) assay, CAR-T cells were mixed with target cells (104 cells) at effector-to-target ratios (E:T = 5:1) and then added to the anti-gamma IFN-γ antibody-precoated plates from the human IFN-γ ELISpot assay kit (DKW22-1000-096s; Dakewei), along with a negative control (effector CD8+ T cells alone) or positive control (phytohemagglutinin stimulation). Plates were incubated for 16–20 h in a humidified atmosphere containing 5% CO2 at 37 °C. The ELISpot assays were then performed according to the manufacturer’s instructions. The plates were scanned by the S6 ultra immunoscan reader (Cellular Technology Ltd) and the number of IFN-γ-positive T cells was calculated by ImmunoSpot software (Version 5.1.34) (Cellular Technology Ltd).
Zou F., Lu L., Liu J., Xia B., Zhang W., Hu Q., Liu W., Zhang Y., Lin Y., Jing S., Huang M., Huang B., Liu B, & Zhang H. (2019). Engineered triple inhibitory receptor resistance improves anti-tumor CAR-T cell performance via CD56. Nature Communications, 10, 4109.
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3

SARS-CoV-2 S Peptide Stimulation Assay

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Indicated cells were cultured with SARS-CoV-2 S peptides pool (GenScript) at a concentration of 2 μg/well for 18-24 h. Antigen specific cells of monkeys, BALB/c mice and human PBMCs were detected by Monkey IFN-γ ELISPOT kit, Monkey IL-4 ELISPOT kit, mouse IFN-γ ELISPOT kit, mouse IL-4 ELISPOT kit and Human IFN-γ ELISPOT kit (Dakewe) according to the manufacturer’s protocol. Antigen-specific spots were then counted using an S6 ultra immunoscan reader (Cellular Technology Ltd.), and the number of IFN-γ- or IL-4-positive T cells was calculated by ImmunoSpot 5.1.34 software (Cellular Technology Ltd.). The number of spots was converted into the number of spots per million cells and plotted as mean ± SEM.
Ma X., Zou F., Yu F., Li R., Yuan Y., Zhang Y., Zhang X., Deng J., Chen T., Song Z., Qiao Y., Zhan Y., Liu J., Zhang J., Zhang X., Peng Z., Li Y., Lin Y., Liang L., Wang G., Chen Y., Chen Q., Pan T., He X, & Zhang H. (2020). Nanoparticle Vaccines Based on the Receptor Binding Domain (RBD) and Heptad Repeat (HR) of SARS-CoV-2 Elicit Robust Protective Immune Responses. Immunity, 53(6), 1315-1330.e9.
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4

SARS-CoV-2 RBD Protein Binding Assay

Cited 1 time
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The detailed protocol was developed by following previous procedures with minor modifications (29 (link)). Briefly, multiScreenHTS IP filter plates of 0.45 μm (Millipore Sigma) were coated with 2.5 mg/mL SARS-CoV-2 RBD protein at 4 °C for 12 hours. Then plates were washed three times with PBS and blocked with DMEM at 37 °C for 1 hour. One million mice bone marrow cells were harvested and incubated inside each well at 37 °C overnight. After washing with PBS/T three times, plates were incubated with anti-IgG-HRP antibody (Jackson ImmunoResearch) at room temperature for 2 hours. Plates were washed with PBS/T three times, RBD-specific spots were then counted using an S6 ultra immunoscan reader (Cellular Technology Ltd.).
Qiao Y., Zhan Y., Zhang Y., Deng J., Chen A., Liu B., Zhang Y., Pan T., Zhang W., Zhang H, & He X. (2022). Pam2CSK4-adjuvanted SARS-CoV-2 RBD nanoparticle vaccine induces robust humoral and cellular immune responses. Frontiers in Immunology, 13, 992062.
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5

NK Cell Cytotoxicity Assay Using IFN-γ ELISpot

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For this assay, NK cells (105 cells) were mixed with target cells (tumor cells or myeloid cells) at effector-to-target (E:T) ratios of 2:1. This mix was then added to anti-IFN-γ pre-coated plates from the human IFN-γ ELISpot assay kit (DKW22-1000-096s; Dakewei, China), along with the following control groups: no antibody, isotype IgG1 control (MCE, HY-P9900, Newark, NJ, USA), and positive control (phytohemagglutinin stimulation). Plates were incubated for 24 h in a humidified atmosphere containing 5% CO2 at 37 °C. The ELISpot assays were then performed according to the manufacturer's instructions. The plates were scanned with the S6 ultra immunoscan reader (Cellular Technology Ltd.) and the number of IFN-γ-positive T-cells was calculated using ImmunoSpot software (Version 5.1.34, Cellular Technology Ltd.).
Tan J., Liu T., Fan W., Wei J., Zhu B., Liu Y., Liu L., Zhang X., Chen S., Lin H., Zhang Y, & Li J. (2022). Anti-PD-L1 antibody enhances curative effect of cryoablation via antibody-dependent cell-mediated cytotoxicity mediating PD-L1highCD11b+ cells elimination in hepatocellular carcinoma. Acta Pharmaceutica Sinica. B, 13(2), 632-647.
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6

Quantifying CAR-T Cell IFN-γ Production

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For the ELISpot assay, CAR-T cells were mixed with target cells (104 cells) at E:T = 4:1 and then added to the anti- IFN-γ antibody-precoated plates from the human IFN-γ ELISpot assay kit (DKW22-1000-096 s; Dakewei, Shenzhen, China), along with a negative control (effector CD8+ T cells alone) or positive control (phytohemagglutinin [PHA] stimulation). Plates were incubated for 20 h in a humidified atmosphere containing 5% CO2 at 37°C. The ELISpot assays were then performed according to the manufacturer’s instructions. The plates were scanned by the S6 ultra immunoscan reader (Cellular Technology, Shaker Heights, OH, USA), and the number of IFN-γ+ T cells was calculated by ImmunoSpot software (version 5.1.34, Cellular Technology).
Zou F., Tan J., Liu T., Liu B., Tang Y., Zhang H, & Li J. (2021). The CD39+ HBV surface protein-targeted CAR-T and personalized tumor-reactive CD8+ T cells exhibit potent anti-HCC activity. Molecular Therapy, 29(5), 1794-1807.
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7

Quantifying Anti-PD-1 Antibody-Secreting Cells

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For enzyme-linked immunosorbent spot (ELISpot) assay, the PVDF plates were coated with PD-1 protein (2 μg/well, Sino Biological) overnight at 4 °C. Mature B cells, PBs, and LLPCs were sorted via flow cytometry and then added to PD-1 coated plates. Untransduced B cells were used as negative control. Plates were incubated for 16–20 h in a humidified atmosphere containing 5% CO2 at 37 °C. HRP conjugated anti-Human IgG-Fc Fragment (A80–104P, Bethyl) was used for detection of bound antibody. The ELISpot assays were then performed according to the manufacturer’s instructions. The plates were scanned by the S6 ultra immunoscan reader (Cellular Technology Ltd) and the number of α-PD-1 antibody positive cells was calculated by ImmunoSpot software (Version 5.1.34; Cellular Technology Ltd).
Luo B., Zhan Y., Luo M., Dong H., Liu J., Lin Y., Zhang J., Wang G., Verhoeyen E., Zhang Y, & Zhang H. (2020). Engineering of α-PD-1 antibody-expressing long-lived plasma cells by CRISPR/Cas9-mediated targeted gene integration. Cell Death & Disease, 11(11), 973.
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