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Glomax multi jr detection system

Manufactured by Promega
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The GloMax Multi JR detection system is a compact and versatile luminometer designed for a wide range of luminescence-based assays. It measures luminescent signals with high sensitivity and accuracy, providing researchers with a reliable tool for their laboratory needs. The core function of this product is to detect and quantify luminescent signals generated in various biological and biochemical applications.

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Lab products found in correlation

Mts 11c mini tubes, by Corning (3 mentions) Atp bioluminescent somatic cell assay kit, by Merck Group (2 mentions) Dual luciferase assay kit, by Promega (2 mentions) Nano glo hibit lytic detection reagent, by Promega (1 mentions) Nano glo luciferase assay system, by Promega (1 mentions) Renilla luciferase assay system, by Promega (1 mentions) Passive lysis buffer (plb), by Promega (1 mentions) 4 methylumbelliferyl, by Merck Group (1 mentions) Smartspec 3000, by Bio-Rad (1 mentions) μquant microplate spectrophotometer, by Agilent Technologies (1 mentions) Dnaqf, by Merck Group (1 mentions) Blyscan glycosaminoglycan assay, by Biocolor (1 mentions) Dual luciferase assay, by Promega (1 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions) Kinase glo luminescent kit, by Promega (1 mentions)

13 protocols using glomax multi jr detection system

1

Dual-Luciferase Reporter Assay Protocol

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In dual-luciferase reporter assays, cells were co-transfected with the listed constructs. After 24 hrs, cells harvested and lysed in passive lysis buffer. Luciferase activities were analyzed by dual-luciferase reporter assay system using GloMax Multi Jr detection system (Promega). Firefly luciferase activity was normalized to the activity of control renilla luciferase. Data represent the mean ± SD from three experiments.
Liu W., Wang C., Wang S., Zeng K., Wei S., Sun N., Sun G., Wang M., Zou R., Liu W., Lin L., Song H., Jin Z, & Zhao Y. (2021). PRPF6 promotes androgen receptor/androgen receptor-variant 7 actions in castration-resistant prostate cancer cells. International Journal of Biological Sciences, 17(1), 188-203.
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2

Quantification of Intracellular ATP in Epimastigotes

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The intracellular ATP was quantified by using an ATP bioluminescent somatic cell assay kit (Sigma-Aldrich). Briefly, mutant epimastigotes (107 parasites per tube, 0.1 ml) were incubated in a solution containing 100 mM sucrose, 50 mM KCl, and 50 mM Tris–HCl (pH 7.2 adjusted with HCl). Cellular extracts were prepared by mixing 0.1 ml epimastigotes with 0.1 ml somatic cell ATP releasing reagent and the mixture was left on ice for 1 min. Half of the cellular extract (0.1 ml) was transferred to MTS-11C minitubes (Axygen, Union City, CA, USA) containing 0.1 ml ATP assay mix and stirred for 10 s at room temperature. The total amount of light emitted was measured with a GloMax Multi JR detection system (Promega, Madison, WI, USA). Total intracellular ATP concentration per cell number was calculated using a standard ATP curve, prepared, and analyzed in each experiment (Carvalho-Kelly et al., 2020 (link)).
Dick C.F., Rocco-Machado N., Dos-Santos A.L., Carvalho-Kelly L.F., Alcantara C.L., Cunha-E-Silva N.L., Meyer-Fernandes J.R, & Vieyra A. (2022). An Iron Transporter Is Involved in Iron Homeostasis, Energy Metabolism, Oxidative Stress, and Metacyclogenesis in Trypanosoma cruzi. Frontiers in Cellular and Infection Microbiology, 11, 789401.
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3

Interferon Luciferase Reporter Assay

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HEK-293T cells were transfected with ISRE-firefly luciferase reporter plasmid, pRL-TK (renilla luciferase) and the indicated expression plasmids. After 24 h transfection, cells were treated with IFN-β and IFN-α (500 IU/mL) for 16 h, and luciferase activity was measured. The dual luciferase assay kit (Promega E1960) was used to perform luciferase assays, and GloMax-Multi JR detection system (Promega) was used to quantify luciferase activity.
Gao X., Tian H., Zhu K., Li Q., Hao W., Wang L., Qin B., Deng H, & Cui S. (2022). Structural basis for Sarbecovirus ORF6 mediated blockage of nucleocytoplasmic transport. Nature Communications, 13, 4782.
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4

Luciferase-Based Bioluminescence Assays

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The activity of renilla luciferase was detected through the renilla luciferase assay system (Promega), following the protocol. HiBiT tag was detected by the Nano-Glo HiBiT Lytic Detection Reagent (Promega). The activity of Nanoluc luciferase was assessed by the Nano-Glo luciferase assay system (Promega) according to the manufacturers’ instructions. Moreover, the GloMax-Multi Jr detection system (Promega) was used to detect bioluminescence.
Wei X.F., Fan S.Y., Wang Y.W., Li S., Long S.Y., Gan C.Y., Li J., Sun Y.X., Guo L., Wang P.Y., Yang X., Wang J.L., Cui J., Zhang W.L., Huang A.L, & Hu J.L. (2022). Identification of STAU1 as a regulator of HBV replication by TurboID-based proximity labeling. iScience, 25(6), 104416.
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5

Caspase-3/7 Activity Assay in Macrophages

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Caspase 3/7 activities in macrophages were measured to determine the apoptotic events, using a Caspase-Glo assay kit. Briefly, the luminogenic substrate containing the tetrapeptide sequence DEVD is cleaved by caspase 3/7. After caspase cleavage, a luciferase substrate is released, resulting in the luciferase reaction and the production of luminescent signal. Cell suspension (200 μl; 1 × 105 cells/ml) was seeded into a 96-well plate and incubated with or without test reagents at 37 °C in DMEM with 5% FBS. After that, an equal volume of reagents was added to each well. Then, samples were incubated at room temperature for 2 h. Finally, the luminescence of each sample was measured by a luminometer (GloMax Multi Jr Detection System, Promega, USA). These fluorescent data from each group were proportional to the amount of caspase activity, which was assigned as the relative fluorescence unit (RFU).
Hu F., Xing F., Zhu G., Xu G., Li C., Qu J., Lee I, & Pan L. (2015). Rhein antagonizes P2X7 receptor in rat peritoneal macrophages. Scientific Reports, 5, 14012.
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6

Quantifying Intracellular ATP in MDA-MB-231 Cells

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The intracellular ATP pool was quantified in the MDA-MB-231 cells using the adenosine 5′-triphosphate (ATP) bioluminescent somatic cell assay kit (Sigma Chemical Co, St. Louis, MO, USA). MDA-MB-231 cells (5 × 104 cells/well) plated in 24-wells were prepared by adding 0.1 mL of somatic cell ATP and releasing reagent in 24-well plates placed ice for 10 min. After that, the cellular extracts were transferred to MTS-11C mini tubes (Axygen Scientific, Union City, CA, USA) containing 0.1 mL of ATP assay mix and mixed for 10 s. The total amount of light emitted was measured with a GloMax Multi JR detection system (Promega Corporation, Fitchburg, WI, USA). The total intracellular ATP concentration per cell number was calculated using a standard ATP curve prepared and analysed in each experiment.
Lacerda-Abreu M.A., Russo-Abrahão T., Rocco-Machado N., Cosentino-Gomes D., Dick C.F., Carvalho-Kelly L.F., Cunha Nascimento M.T., Rocha-Vieira T.C, & Meyer-Fernandes J.R. (2021). Hydrogen Peroxide Generation as an Underlying Response to High Extracellular Inorganic Phosphate (Pi) in Breast Cancer Cells. International Journal of Molecular Sciences, 22(18), 10096.
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7

Dual Luciferase Assay for IFN-β Signaling

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Firefly luciferase IFNβ reporter plasmid (100 ng) and pRL-TK (Renilla luciferase) plasmid (internal control) (10 ng) were transfected together with the indicated expression plasmids (total 800 ng) into HEK293 cells at a density of 3 × 105 cells per well of 12-well plates. If cotransfected with MAVS or STING expression plasmid, at 24 hours after transfection, then cells were lysed by passive lysis buffer (Promega E1941) for the following luciferase assays. In other cases, transfected cells were cultured for 24 hours, then challenged by transfection of poly(I:C) (1 μg/ml) and poly(dA:dT) (1 μg/ml), and lysed by passive lysis buffer at 18 hours after treatment. The dual luciferase assay kit (Promega E1960) was used to perform luciferase assays, and GloMax-Multi Jr. detection system (Promega) was used to quantify luciferase activity. The relative luciferase activity was calculated by normalizing firefly to Renilla luciferase activity.
Yu K., Tian H, & Deng H. (2020). PPM1G restricts innate immune signaling mediated by STING and MAVS and is hijacked by KSHV for immune evasion. Science Advances, 6(47), eabd0276.
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8

Chitinolytic Activity Assay of B. thuringiensis

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To test the chitinolytic activity of B. thuringiensis strains, ~1 × 108 cells mL−1 were used to inoculate nutrient broth (DB Bioxon), and cultures were grown at 28°C, 180 rpm for ~72 h to reach autolysis. Samples were collected in duplicate to determine the optical density at 600 nm, using a Smart Spec3000 (BioRad, Hercules, CA). Three fluorogenic chitin derivatives, 4‐methylumbelliferyl‐β‐D‐N,N′,N′′‐triacetylchitotriose [4‐MU‐(GlcNAc)3] (tetrameric fluorescent derivative), 4‐methylumbelliferyl‐β‐D‐N,N′‐diacetylchitobioside [4‐MU‐(GlcNAc)2] (trimeric fluorescent derivative), and 4‐methylumbelliferyl‐N‐acetyl‐β‐D‐glucosaminide [4‐MU‐GlcNAc] (dimeric fluorescent derivative) (Sigma, St. Louis, MO) were used to evaluate endochitinase, chitobiosidase, and N‐acetylglucosaminidase activities, respectively, in a 100 mmol L−1 phosphate reaction buffer (pH 7.0) (Barboza‐Corona et al. 2003). 4‐MU released from the fluorogenic substrate was calculated fluorometrically with excitation at 360 nm and emission at 455 nm (Glomax Multi Jr. Detection System, Promega, Sunnyvale, CA), using a 4‐MU standard curve. One unit (U) of chitinolytic activity was defined as the amount of enzyme required to release 1 μmol of 4‐methylumbelliferone in 1 h (Barboza‐Corona et al. 2003).
de la Fuente‐Salcido N.M., Casados‐Vázquez L.E., García‐Pérez A.P., Barboza‐Pérez U.E., Bideshi D.K., Salcedo‐Hernández R., García‐ Almendarez B.E, & Barboza‐Corona J.E. (2016). The endochitinase ChiA Btt of Bacillus thuringiensis subsp. tenebrionis DSM‐2803 and its potential use to control the phytopathogen Colletotrichum gloeosporioides. MicrobiologyOpen, 5(5), 819-829.
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9

Quantification of Cartilage Composition

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For measurement of DNA and characterization of the cartilage extracellular matrix (n = 12 per group), cartilage and bone were separated, freeze dried, and digested in 100 mM sodium phosphate buffer/10 mM Na2EDTA/10 mM L-cysteine/0.125 mg/mL papain overnight at 60 °C. DNA was analyzed by fluorescence assay using bisBenzimide (DNAQF, Sigma). Fluorescence was read using a Glomax Multi Jr Detection System (Promega, Madison, WI). DNA content was quantified by comparison to a standard curve generated using known amounts of calf thymus DNA. Cartilage GAG content was determined using the Blyscan Glycosaminoglycan Assay based on binding to dimethylmethylene blue dye (Biocolor, Carrickfergus, County Antrim, UK). Precise quantities were determined from a standard curve developed using the chondroitin sulphate supplied with the kit. Cartilage collagen content was determined using the chloramine-T hydroxyproline assay according to Reddy and Enwemeka [19 (link)]. Colorimetric results were obtained using a μQuant Microplate Spectrophotometer (Biotek, Winooski, VT). Hydroxyproline was used to develop a standard curve, and the amount of collagen was calculated assuming 12.5% of collagen is hydroxyproline. DNA, GAG, and collagen contents were normalized to tissue dry weight.
Elder S., Chenault H., Gloth P., Webb K., Recinos R., Wright E., Moran D., Butler J., Borazjani A, & Cooley A. (2018). Effects of Antigen Removal on a Porcine Osteochondral Xenograft for Articular Cartilage Repair. Journal of biomedical materials research. Part A, 106(8), 2251-2260.
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10

Keap1/Nrf2 Regulation via Pten in MEF

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Keap1 & Nrf2 double-disrupted MEF were seeded at 5 × 105 cells/well in 6-well plates on day 0. Plasmid DNA (5 ng pCMV Nrf2, 10 ng pRLTK- ΔARE, 100 ng pCMV NQO1-ARE-Luc, 2.5 ng pCMV WT Keap1) was transfected with Lipofectamine 2000 (Invitrogen, Carlsbad, CA) on day 1. pCMV mock vector was utilized to maintain total DNA transfected at 400 ng. Cells were treated with inducers (3 μM WA, 10 μM sulforaphane, 25 nM CDDO-Im) 24 hours after transfection (day 2). For Pten silencing or overexpression experiments, scrambled/siPTEN (SC-36326, Santa Cruz, Dallas, TX) or wild-type PTEN (500 ng) constructs were transfected 24 hours prior to transfection with Nrf2 expression plasmid. Dual Luciferase Assay (Promega, Madison, WI) was carried out 24 hours later (day 3). Luminescence was measured using a Glomax Multi Jr Detection system (Promega, Madison, WI).
Palliyaguru D.L., Chartoumpekis D.V., Wakabayashi N., Skoko J.J., Yagishita Y., Singh S.V, & Kensler T.W. (2016). Withaferin A induces Nrf2-dependent protection against liver injury: role of Keap1-independent mechanisms. Free radical biology & medicine, 101, 116-128.
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