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Glomax explorer plate reader

Manufactured by Promega
Sourced in United States

The GloMax Explorer is a versatile multimode plate reader that can perform a wide range of luminescent, fluorescent, and absorbance-based assays. It is equipped with high-performance optics and an advanced detection system to enable sensitive and accurate measurements across various applications in life science research and drug discovery.

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24 protocols using glomax explorer plate reader

1

Cas13 Activity Assay Optimization

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The reaction mix for Cas13 activity was prepared by combining 8.6 μl of nuclease-free water, 2 μl of cleavage buffer (400 mM Tris pH 7.4), 2 μl of LwaCas13a protein diluted in Storage Buffer (SB) to a concentration of 126.6 μg/ml, 1 μl of crRNA (40 ng/μl), 1 μl of the fluorescent reporter (IDT, diluted in water to a final concentration of 4 μM), 1 μl of Murine RNase inhibitor (NEB, #M0314), 0.8 μl of rNTP solution mix (25 mM each, NEB, #N0466), 0.6 μl of NxGen T7 RNA Polymerase (Lucigen, #30223-2) and 1 μl of MgCl2 (120 mM). 2 μl of the RT-RPA-amplified product were then added to the mix. The 20μl-LwaCas13a reactions were transferred in 5μl-replicates (4 wells each sample) to a 384-well, round, black-well, clear-bottom plate (Corning, #3544). The plate was briefly spun down at 500 g for 15 sec to remove potential bubbles and placed into a preheated GloMax® Explorer plate reader (Promega) at 37 °C. Fluorescence was measured every 5 min for 3 h. Data analysis, if not otherwise stated, was performed at the 30-min time-point. The reporter sequence is provided in Supplementary Table 6.
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2

ERα Binding Assay Using EFC Method

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Competition binding assays were performed by using an enzyme fragment complementation (EFC) method described in the HitHunter (Freemont, CA) EFC Estrogen Chemiluminescence Assay kit according to the manufacturer’s instructions. Briefly, competing ligands at final concentrations ranging from 25 pM to 2 µM were incubated with 5 nM recombinant ERα (Invitrogen) and 17β-estradiol-conjugated enzyme donor for 1.5 h. The enzyme acceptor was then added followed by the chemiluminescence substrate and incubated for 1 h. Relative luminescence was determined by using a GloMax Explorer plate reader (Promega). Sigmoidal standard curves were created by Excel23 (link).
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3

Myeloma Cell-Stromal Cell Coculture Assay

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Cocultures were seeded into 384 well plates (Corning) with the Multidrop Combi (Thermo Scientific). 800 stromal cells were seeded and incubated overnight. 17 hours later, 700 myeloma cells were added on top of the stromal cells. On the third day, 24 hours after the addition of myeloma cells, cocultures were treated with tofacitinib (LC Laboratories), ruxolitinib (Selleck Chemicals), JAK3i,20 (link) or IL-6 blocking antibody (R&D Systems). For drug combination studies, on the fourth day, melphalan (Sigma Aldrich), carfilzomib (Selleck Chemicals), or venetoclax (Selleck Chemicals) were additionally added to cocultures. On the fifth day, myeloma cell viability was detected with the addition of luciferin (Gold Biotechnology) and read for luminescence on Glomax Explorer plate reader (Promega) as previously described.21 (link) For monoculture studies cell viability was measured using CellTiter-Glo reagent (Promega). All measurements were performed in quadruplicate. All viability data are reported as normalized to dimethyl sulfoxide (DMSO)-treated cell line in monoculture.
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4

In Vitro Translation Assay

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5 μl in vitro translation reactions were set up with 2.5 μl of lysate and 20 ng total RNA (0.84 mM ATP, 0.21 mM GTP, 21 mM creatine phosphate, 0.009 units/ml creatine phosphokinase, 10 mM HEPES pH 7.5, 2 mM DTT, 2 mM MgOAc, 100 mM KOAc, 0.008 mM amino acids, 0.25 mM spermidine, 5 units RNasin Plus RNase inhibitor (Promega) as described (48 (link)). Reaction tubes were incubated at 30°C for 45 min, and expression of the reporter was measured using the Renilla Luciferase Assay System (Promega) on a GloMax Explorer plate reader (Promega).
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5

Quantifying Cellular ATP Levels

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ATP measurement was done using CellTiter-Glo Luminescent Cell Viability Assay (Promega Madison, WI USA) following manufacturer instructions. Measuarements are made using the GloMax explorer plate reader (Promega Madison, WI USA).
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6

Brilacidin Cytotoxicity Evaluation in Vero and HSAEC Cells

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Vero cells and HSAECs were seeded in 96-well white plates at 5 × 104 cells per well, respectively, and allowed to grow for 24 h. Brilacidin was diluted to the desired concentrations in appropriate cell culture media. Dilutions of brilacidin were applied to individual wells of the plate and incubated at 37 °C with 5% CO2 for 24 h. Brilacidin and media mixture were removed, and cell viability was measured with CellTiter-Glo® Luminescent Cell Viability Assay per manufacturer’s instructions (Promega, G7572, Madison, WI, USA). Luminescence was measured using GloMax Explorer Plate Reader (Promega, GM3510, Madison, WI, USA).
Viral infections were conducted as previously outlined in 96-well white plates. At specified collection time, CellTiter-Glo® Luminescent Cell Viability Assays were conducted according to manufacturer’s protocol. Percent cell survival was analyzed and compared with mock infected wells.
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7

Assessing Cell Viability after HIV-1 Treatment

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Cell viability was assessed to confirm survival rates of HIV-1-infected and uninfected cells after treatment with IR in combination with Rapa or INK128. A concentration of 5 × 104 cells in 100 uL was plated in triplicate on a 96-well plate. After plating and treatment, cells were incubated for 2, 3, or 5 days (according to the therapeutic regimen), and cell viability assessed with 100 uL of Cell-Titer Glo reagent (Cat. #: G7572; Promega, Madison, WI, USA). A GloMax explorer plate reader (Promega) was used to measure relative luminescence units (RLU) resulting from ATP levels in cultured cells.
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8

In Vitro Translation Assay

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A total of 5 µL in vitro translation reactions were set up with 2.5 µL of lysate and 20 ng total RNA (0.84 mM ATP, 0.21 mM GTP, 21 mM Creatine Phosphate, 0.009 units/mL Creatine phosphokinase, 10 mM HEPES pH 7.5, 2 mM DTT, 2 mM MgOAc, 100 mM KOAc, 0.008 mM amino acids, 0.25 mM spermidine, 5 units RNasin Plus RNase inhibitor [Promega]) as described by Lee et al. (2015) (link). Reaction tubes were incubated at 30°C for 45 min, and expression of the reporter was measured using the Renilla Luciferase Assay System (Promega) on a GloMax Explorer Plate Reader (Promega).
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9

Organoid Viability and Apoptosis Assay

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Viability and apoptosis of basal‐out and apical‐out organoids were assessed using the RealTime‐Glo MT Cell Viability Assay (Promega) and RealTime‐Glo Annexin V Apoptosis Assay (Promega; referred to as ‘apoptosis and necrosis assay’). Equal numbers of organoids were seeded into each well of a white 96‐well plate with clear bottom to induce polarity reversal as described above. Detection reagents were prepared according to the manufacturer's instructions and added to the respective wells. Luminescence was measured at 0, 12, 24, 36, 48, 60 and 72 h after adding the substrates at time point 0 h using a GloMax Explorer plate reader (Promega). Experiments were carried out in eight technical replicates of three biological replicates per intestinal section (small and large intestines).
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10

In Vitro Translation Assay

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5 μL in vitro translation reactions were set up with 2.5 μL of lysate and 20 ng total RNA (0.84mM ATP, 0.21mM GTP, 21mM Creatine Phosphate, 0.009units/mL Creatine phosphokinase, 10mM HEPES pH 7.5, 2mM DTT, 2mM MgOAc, 100mM KOAc, 0.008mM amino acids, 0.25mM spermidine, 5 units RNasin Plus RNase inhibitor (Promega) as described (Lee et al. 2015). Reaction tubes were incubated at 37°C for 45 minutes, and expression of the reporter was measured using the Renilla Luciferase Assay System (Promega) on a GloMax Explorer plate reader (Promega).
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