Human immunology v2 panel

Manufactured by NanoString

The Human Immunology V2 panel is a comprehensive gene expression profiling tool designed to analyze the immune system. It targets over 700 genes related to various aspects of human immunology, including innate and adaptive immune responses, immunoregulation, and cellular interactions. The panel provides a detailed snapshot of the immune landscape within a sample, enabling researchers to gain deeper insights into immune system function and dynamics.

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Lab products found in correlation

Anti cd19 magentic beads, by Miltenyi Biotec (2 mentions) Ncounter, by NanoString (2 mentions) Ncounter low rna input amplification kit, by NanoString (1 mentions) M mlv reverse transcriptase enzyme, by Thermo Fisher Scientific (1 mentions) Ncounter platform, by NanoString (1 mentions)

4 protocols using human immunology v2 panel

Nanostring-Based Analysis of EV-Derived Transcripts

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The nCounter Low RNA Input Amplification Kit (NanoString Technologies, Seattle, WA) was used to retrotranscribe and pre-amplify 4 μL EV-derived RNA using 10 cycles. Retrotranscription was carried out in 0.5 mL tubes while pre-amplification, using primers targeting the genes of the Human Immunology V2 Panel (NanoString Technologies), was performed in 384-well plates to prevent sample evaporation. In parallel, a Moloney Murine Leukema Virus (M-MLV) Reverse Transcriptase Enzyme (Thermo Fisher Scientific) was also tested for cDNA synthesis. The Human Immunology V2 Panel (NanoString Technologies) was used to analyze EV-derived, pre-amplified cDNA according to manufacturer instructions. This panel targets 594 general genes involved in the immune response such as cytokines, enzymes, interferons and their receptors. Samples were hybridized for 18 h at 65 °C.

Bracht J.W., Gimenez-Capitan A., Huang C.Y., Potie N., Pedraz-Valdunciel C., Warren S., Rosell R, & Molina-Vila M.A. (2021). Analysis of extracellular vesicle mRNA derived from plasma using the nCounter platform. Scientific Reports, 11, 3712.

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RNA Expression Profiling of RH30_19 Cells

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RNA was extracted from in vitro cultures of RH30_19 cells or from single cell suspensions of tumor tissue. RNA was isolated from 3,000 cells/μL per sample. RH30_19 tumor cells were isolated from whole-tumor samples using anti-CD19 magentic beads (Miltenyi Biotec). RNA expression profiling was performed with the Nanostring nCounter using the Human Immunology V2 panel (Nanostring Technologies). Normalization and analysis was conducted in Rosalind (Nanostring Technologies). Gene categories were selected in Microsoft Excel using the Human Immunology V2 panel Annotations summary.

Sullivan P.M., Kumar R., Li W., Hoglund V., Wang L., Zhang Y., Shi M., Beak D., Cheuk A., Jensen M.C., Khan J., Dimitrov D.S, & Orentas R.J. (2022). FGFR4-Targeted Chimeric Antigen Receptors Combined with Anti-Myeloid Polypharmacy Effectively Treat Orthotopic Rhabdomyosarcoma. Molecular Cancer Therapeutics, 21(10), 1608-1621.

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Immunological Profiling of RH30_19 Cells

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RNA was extracted from in vitro cultures of RH30_19 cells or from single cell suspensions of tumor tissue. RNA was isolated from 3000 cells/ul per sample. RH30_19 tumor cells were isolated from whole tumor samples using anti-CD19 magentic beads (Miltenyi Biotec). RNA expression profiling was performed with the Nanostring nCounter using the Human Immunology V2 panel (Nanostring Technologies). Normalization and analysis was conducted in Rosalind (Nanostring Technologies). Gene categories were selected in Microsoft Excel using the Human Immunology V2 panel Annotations summary.

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Immunology Gene Expression Profiling

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FFPE- or cell pellet-extracted RNA (approximately 100 ng) was analyzed using the human Immunology V2 panel and the nCounter platform (NanoString Technologies). This panel profiles 594 immunology-related human genes as well as two types of built-in controls: positive controls (spiked RNA at various concentrations to evaluate the overall assay performance) and 15 housekeeping genes (to normalize for differences in total RNA input). Sample preparation and hybridization was carried out according to the manufacturer’s instructions. In brief, 5 μL (100 ng) of total RNA was hybridized at 96°C overnight and then digitally analyzed for frequency of each RNA species. Data were collected using the nCounter Digital Analyzer, and data normalization and analysis were carried out using the nSolver software (V.3). Normalization factors were derived from the geometric mean of housekeeping genes, mean of negative controls, and geometric mean of positive controls.

Bhatia S., Longino N.V., Miller N.J., Kulikauskas R., Iyer J.G., Ibrani D., Blom A., Byrd D.R., Parvathaneni U., Twitty C.G., Campbell J.S., Le M.H., Gargosky S., Pierce R.H., Heller R., Daud A.I, & Nghiem P. (2019). Intratumoral Delivery of Plasmid IL12 Via Electroporation Leads to Regression of Injected and Noninjected Tumors in Merkel Cell Carcinoma. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(3), 598-607.

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