Quantigene plex 2

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Quantigene Plex 2.0 is a multiplex gene expression quantification system designed for accurate and sensitive measurement of gene expression levels across multiple targets. The system utilizes a branched DNA signal amplification technology to provide quantitative results.

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Lab products found in correlation

Glomax multi, by Promega (2 mentions) Paxgene rna tube, by Qiagen (2 mentions) Panel 11834, by Thermo Fisher Scientific (2 mentions) β actin, by Thermo Fisher Scientific (1 mentions) Luminex instrument, by Bio-Rad (1 mentions) Xn 9000, by Sysmex (1 mentions)

Close spelling variants of this product

QuantiGene ViewRNA ISH Tissue-2 Plex Assay Quantigene Plex 2.0 reagent system QuantiGene 2.0 Plex Assay kit QuantiGene Plex 2.0 Assay QuantiGene ViewRNA ISH Tissue 2-Plex Assay kit QuantiGene Plex

5 protocols using quantigene plex 2

Measuring A3AR Expression in Blood

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The peripheral blood expression of A3AR was measured at baseline and Week 12. A3AR mRNA expression in white blood cells was determined from blood collected to PAXgene RNA tubes (Qiagen), using the QuantiGene Plex 2.0^® assay (Thermo Fisher). β‐actin was used as a reference control, and the oligonucleotide probe sets were designed by Thermo Fisher. Luminescence from each specific probe set was captured by Glomax Multi (Promega). A3AR was expressed in units, where one unit was defined as the mean of A3AR expression in healthy subjects, as previously determined.¹⁶

Safadi R., Braun M., Francis A., Milgrom Y., Massarwa M., Hakimian D., Hazou W., Issachar A., Harpaz Z., Farbstein M., Itzhak I., Lev‐Cohain N., Bareket‐Samish A., Silverman M.H, & Fishman P. (2021). Randomised clinical trial: A phase 2 double‐blind study of namodenoson in non‐alcoholic fatty liver disease and steatohepatitis. Alimentary Pharmacology & Therapeutics, 54(11-12), 1405-1415.

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A3AR Expression in WBC and HCC Response

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Another secondary objective was to evaluate the relationship between white blood cell (WBC) A3AR expression (which has been suggested to mirror the expression in HCC tumor cells [6 (link),7 (link),29 (link),30 (link)]) as assessed at baseline and every cycle thereafter at selected study centers, (n = 53 patients) and clinical response. A3AR mRNA expression in WBC was determined from blood collected to a PAXgene RNA tube (Qiagen, Venlo, The Netherlands), using the QuantiGene Plex 2.0^® assay (Thermo Fisher, Waltham, MA, USA). β-actin was used as a reference control, and the oligonucleotide probe sets were designed by Thermo Fisher. Luminescence from each specific probe set was captured by GloMax Multi (Promega, Madison, WI, USA). A3AR was expressed in units, where 1 unit was defined as the mean of A3AR expression in healthy subjects (n = 50). Healthy subjects were 20–70 years of age with no known illness and no prior treatment.

Stemmer S.M., Manojlovic N.S., Marinca M.V., Petrov P., Cherciu N., Ganea D., Ciuleanu T.E., Pusca I.A., Beg M.S., Purcell W.T., Croitoru A.E., Ilieva R.N., Natošević S., Nita A.L., Kalev D.N., Harpaz Z., Farbstein M., Silverman M.H., Bristol D., Itzhak I, & Fishman P. (2021). Namodenoson in Advanced Hepatocellular Carcinoma and Child–Pugh B Cirrhosis: Randomized Placebo-Controlled Clinical Trial. Cancers, 13(2), 187.

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mRNA Quantification Using QuantiGene Plex 2.0

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Messenger RNA (mRNA) was quantified using Quantigene Plex 2.0 technology according to manufacturer’s protocol (Affymetrix, Inc., Santa Clara, CA) and as previously described [12 (link)]. Probe sets were designed against the human genome for analysis of Nrf2 target genes, and a modified version of Panel 11834 (Affymetrix) was used. Human primary keratinocyte data were normalized to the housekeeping gene PPIB. Human skin explant data were normalized to the average of housekeeping genes RPL13A and PPIB.

Reisman S.A., Goldsberry A.R., Lee C.Y., O’Grady M.L., Proksch J.W., Ward K.W, & Meyer C.J. (2015). Topical application of RTA 408 lotion activates Nrf2 in human skin and is well-tolerated by healthy human volunteers. BMC Dermatology, 15, 10.

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Quantifying TTK mRNA in HCC samples

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The mRNA expression of TTK in HCC specimens and their adjacent non-tumourous liver tissues were quantified using QuantiGene Plex 2.0^® assay (Affymetrix) following the manufacturer's instructions. All the oligo nucleotide probe sets including capture, label and blocker probes were designed by the manufacturer. For each sample, 200 μg extracted total RNA was used at the beginning of the assay. Mean fluorescence intensity (MFI) generated from each specific probe set were captured and quantified using a Luminex instrument (Bio-Rad).

Liu X., Liao W., Yuan Q., Ou Y, & Huang J. (2015). TTK activates Akt and promotes proliferation and migration of hepatocellular carcinoma cells. Oncotarget, 6(33), 34309-34320.

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Quantifying PBMCs and Nrf2 Transcripts

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PBMCs were isolated from blood collected at each timepoint. Day 28 collections were missing for the 15 mg cohort due to patient refusing blood draw (1), and due to missed visit (1). mRNA was quantified using Quantigene Plex 2.0 technology according to manufacturer’s protocol (Affymetrix Inc., Santa Clara, CA, USA) and as previously described.13 (link) Probe sets were designed against the human genome for analysis of Nrf2 target genes; a modified version of Panel 11834 (Affymetrix) was used. Genes were normalized to the mean expression of housekeeping genes RPL13A (60S ribosomal protein L13a) and peptidyl-prolyl cis–trans isomerase B.
Peripheral blood cells were quantified by automated cell counter using flow cytometry for white blood cell differential (Sysmex, XN9000).

Creelan B.C., Gabrilovich D.I., Gray J.E., Williams C.C., Tanvetyanon T., Haura E.B., Weber J.S., Gibney G.T., Markowitz J., Proksch J.W., Reisman S.A., McKee M.D., Chin M.P., Meyer C.J, & Antonia S.J. (2017). Safety, pharmacokinetics, and pharmacodynamics of oral omaveloxolone (RTA 408), a synthetic triterpenoid, in a first-in-human trial of patients with advanced solid tumors. OncoTargets and therapy, 10, 4239-4250.

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