Establishment of human keratinocyte HaCaT-ARE-GFP-luciferase cells were previously described [17 (link)]. In brief, HaCaT-ARE-luciferase cells were seeded in six-well plates, cultured until 70% confluence, and exposed to ethanol extract of Chaenomeles sinensis. After treatment, cells were lysed with a luciferase lysis buffer (0.1 M potassium phosphate buffer at pH 7.8, 1% Triton X-100, 1 mM DTT, 2 mM EDTA) and the resulting luciferase activity was measured by the GLOMAX Multi-system (Promega, Madison, WI, USA). The data is depicted as a fold ratio of the firefly luciferase activity compared with the control after normalization with the protein concentration.
Glomax multi system
Manufactured by Promega
Sourced in United States
The GLOMAX Multi-system is a versatile luminescence detection platform designed for a wide range of life science applications. It offers a comprehensive solution for measuring various luminescence-based assays, including luciferase reporter gene assays, ATP-based cell viability assays, and bioluminescence resonance energy transfer (BRET) experiments. The GLOMAX Multi-system provides researchers with a reliable and highly sensitive tool to quantify luminescent signals accurately and efficiently.
Automatically generated - may contain errors
Lab products found in correlation
Quantifluor dsdna kit, by Promega (2 mentions)
Miseq platform, by Illumina (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Inorganic pyrophosphatase, by New England Biolabs (1 mentions)
Recombinant rnase inhibitor, by New England Biolabs (1 mentions)
Sp6 polymerase, by New England Biolabs (1 mentions)
Evos fl auto imaging system, by Thermo Fisher Scientific (1 mentions)
Passive lysis buffer (plb), by Promega (1 mentions)
Nad nadh assay kit, by Abcam (1 mentions)
13 protocols using glomax multi system
1
Measuring Antioxidant Activity in HaCaT Cells
2
Renilla Luciferase Assay in Nicotiana benthamiana
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The RLuc-PepMV-3UTR plasmid was obtained by insertion of a DNA fragment covering the PepMV 3′UTR (Baseclear, Leiden, The Netherlands) into a Renilla luciferase reporter plasmid previously described29 (link). Templates for transcription were obtained by PCR using the forward primer SP6FLU and reverse primer (pep1, pA2, or pA6—Supplementary Table S1 ). Transcription reactions were carried out as described above, but with the kit enzyme mix substituted with a 7:2:1 mixture of SP6 polymerase, recombinant RNAse inhibitor, and inorganic pyrophosphatase (all New England Biolabs). 5 × 105 N. benthamiana protoplasts were transfected with 3 µg of RNA and after 16 h incubation at 25 °C under constant light, were freeze-dried and sent by airmail for analysis at Leiden University. Upon arrival material was resuspended in 100ul Tris (10 mM pH 8) and luciferase activity in 50 µl samples was measured using a GloMax multi system (Promega).
3
Transcriptional Regulation Assay in A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549 and A549-FLAG-BAP1 cells (2.0 × 106 cells/well) were cotransfected with pGL3-ARE-luciferase and pGL4.74 vectors. After 24 h transfection, cells were washed three times with 1× PBS and lyzed with luciferase lysis buffer (0.1 M potassium phosphate buffer at pH 7.8, 1% Triton X-100, 1 mM DTT, 2 mM EDTA) for 1 h. The firefly luciferase activity monitored by GLOMAX Multi-system (Promega, Madison, WI, USA) and normalized by the renilla luciferase activity.
4
Firefly Luciferase Reporter Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
B16F10-M-box/E-box-GFP-luciferase cells and B16F10-ARE-GFP-luciferase cells were seeded at a density of 1 × 105 cells in 24-well culture plates. After treatment, B16F10-M-box/E-box-GFP-luciferase cells and B16F10-ARE-GFP-luciferase cells were washed with 1x PBS and lyzed with luciferase lysis buffer [0.1 M potassium phosphate buffer at pH 7.8, 1% Triton X-100, 1 mM DTT, 2 mM EDTA] for 1 h. The resulting firefly luciferase activity was monitored by GLOMAX Multi-system (Promega, Madison, WI, USA), and the luciferase activity was normalized by protein concentration of lysates.
5
Microbial DNA Extraction from Bamboo and Fecal Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA extraction for microbial community analysis was conducted on 1 g bamboo material, 1 g of fecal sample, or on the pellet resulting from 2 mL of either full inocula or de-H2O reactors. The DNA was extracted with phenol-chloroform and precipitated with ice-cold isopropyl alcohol and 3M sodium acetate. DNA pellets were dried and resuspended in TE buffer and stored at −20°C. DNA quality was assessed using 1% (w:v) agarose gel electrophoresis (Life technologiesTM, ES), and quantified by a fluorescence assay (QuantiFluor® dsDNA kit; Promega, United States) using a Glomax®-Multi + system (Promega). Samples were normalized to 1 ng DNA μL–1 and sent to LGC Genomics (DE) for library preparation and sequencing via an Illumina Miseq platform (see Supplementary Information ).
6
Quantifying AP-1 Transcriptional Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
pGreenFire-AP-1-GFP-luciferase cells were seeded at a density of 1×105 cells in 24-well plates. After treatment, pGreenFire-AP1-GFP-luciferase cells were washed with 1× PBS and lysed with luciferase lysis buffer [0.1 M potassium phosphate buffer at pH 7.8, 1% Triton X-100, 1 mM DTT, and 2 mM EDTA] for 1 h. The resulting firefly luciferase activity was monitored using the GLOMAX Multi-system (Promega, Madison, WI, USA). Luciferase activity was normalized to the protein concentration of the lysates.
7
Microbial Community DNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA extraction for microbial community analysis was conducted as in [32 (link)]. Briefly, 1 g of fecal sample or the pellet resulting from 2 mL of liquid sample from fermentation vessels was used. The DNA was extracted with phenol–chloroform and precipitated with ice-cold isopropyl alcohol and sodium acetate. DNA pellets were dried and resuspended in TE buffer. DNA quality was assessed using agarose gel electrophoresis (Life technologiesTM, Waltham, MA, USA), and quantified by a fluorescence assay (QuantiFluor® dsDNA kit; Promega, USA) using a Glomax®-Multi+ system (Promega, Madison, WI, USA). Samples were normalized to 1 ng DNA µL−1 and sent to LGC Genomics (DE) for library preparation and sequencing via an Illumina Miseq platform (see Supplementary Information ).
8
Transfection of BHK21J Cells for Luciferase Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 100 ng of transcript (as judged by gel electrophoresis) was mixed with Dulbecco's Modified Eagle's Medium (DMEM) and 0.5 µL of MessengerMax lipofectamine (Invitrogen) premixed with DMEM, and after 20 min incubation at RT added to a well of a 48-well plate containing BHK21J cells at ∼70% confluency. Cells were grown at 37°C in 5% CO2 on growth medium consisting of DMEM supplemented with 10% GlutaMAX (Thermo Fisher), ampicillin, streptomycin, and fetal calf serum, and analyzed 20–24 h after transfection. Fluorescent microscopy was performed on an EVOS FL Auto Imaging System (Fisher Scientific). Luciferase activity was measured after lysis of cells using Passive Lysis Buffer (Promega) and transferring the contents to a 96-well plate to which the appropriate substrate was added and assayed using a GloMax Multi System (Promega).
9
Quantification of Cellular NAD+ and NADH Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total amount of nicotinamide adenine dinucleotide (NAD+ and NADH) levels were conducted according to an NAD+/NADH Assay Kit (Abcam, ab176723, Cambridge, UK). Briefly, 25 μL of the sample, blank control, or standard were added to 25 μL of specific control or NAD+ extraction solutions. After 15 min of incubation at 37 °C, another 25 μL of control solution or NADH extraction solution (to neutralize NAD+ extracts) was added. Next, 75 μL of NADH reaction mixture was added to each well. After incubation (30 min at room temperature in the dark), fluorescence was measured at Ex = 525 nm and Em = 580–640 nm in a GloMax-Multi+ System (Promega Corporation; Madison, WI, USA). The amount of NADH was calculated by the formula: total (NAD+ and NADH) minus NAD+. The concentration of NAD+, NADH, and total are expressed as μM.
10
Luciferase Assay for ARE-GFP Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A549-ARE-GFP-luciferase and H1299-ARE-GFP-luciferase cells were previously established in our laboratory (Lee et al., 2018b (link)). After treatment, the cells were washed three times with ice-cold 1x PBS and lysed with luciferase lysis buffer (100 mM potassium phosphate buffer at pH 7.8, 1% Triton X-100, 1 mM DTT, and 2 mM EDTA) for 1 h. The cell lysates were collected by centrifugation and the luciferase activity was monitored using the GLOMAX Multi-system (Promega, Madison, WI, United States) followed by normalization of the protein concentration.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!