Recombinant luciferase

Manufactured by Promega

Sourced in China

Recombinant luciferase is an enzyme produced through genetic engineering. It catalyzes a bioluminescent reaction, resulting in the emission of light. This property makes recombinant luciferase a useful tool in various research and analytical applications.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

QuantiLum Recombinant Luciferase (19 citations) Ultra-Glo™ Recombinant Luciferase (3 citations) Recombinant luciferase protein (2 citations) Recombinant firefly luciferase (2 citations)

14 protocols using recombinant luciferase

Quantifying Luciferase Inhibition Screening

Check if the same lab product or an alternative is used in the 5 most similar protocols

To quantify the rate
of false positives reported during the PbLuc liver stage reconfirmation
screen, hits selected for tertiary round screening (405 compounds
from fresh DMSO stocks) were also tested for luminescent interference
between recombinant firefly luciferase and luciferin. Initially, 40
nL of selected compounds was dispensed using a Gen 4 Plus Acoustic
Transfer System (Biosero, San Diego, USA) at 1:3 in 12-point dose
response format (25 to 141.13 × 10^–6 μM;
final DMSO concentration of 0.5% per well) in 1536-well, white, opaque-bottom
plates (ref# 789173-F, Greiner Bio-One). A 24-point single concentration
series of Luciferase Inhibitor-II (9.8 μM per well) was dispensed
as a positive control, while 96 wells of DMSO at 0.5% per well acted
as the negative control. Separately, a solution of 20 pM recombinant
luciferase (Promega cat# E170A) was prepared on ice by successive
dilutions (first 1:999 then into final solution at 20 pM) in phenol
naive DMEM (Invitrogen cat# 31053-028), 5% FBS (Corning), and 5×
Pen Strep Glutamine (Invitrogen). This solution was dispensed at 8
μL per well into the previously spotted plates using a MultiFlo
dispenser (BioTek) and incubated at room temperature for 1 h prior
to the addition of 1 μL of BrightGlo (Promega). Immediately
after the addition of BrightGlo, plates were read using the EnVision
Microplate reader (PerkinElmer).

Abraham M., Gagaring K., Martino M.L., Vanaerschot M., Plouffe D.M., Calla J., Godinez-Macias K.P., Du A.Y., Wree M., Antonova-Koch Y., Eribez K., Luth M.R., Ottilie S., Fidock D.A., McNamara C.W, & Winzeler E.A. (2020). Probing the Open Global Health Chemical Diversity Library for Multistage-Active Starting Points for Next-Generation Antimalarials. ACS Infectious Diseases, 6(4), 613-628.

+ Open protocol

+ Expand

Cell Cytotoxicity Screening of Nanoemulsions

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cell cytotoxicity determinations were performed using an automated luminescence assay based on the luciferin reaction. Recombinant luciferase (Promega Corp., Madison, WI) was used to catalyze the conversion of luciferin substrate to oxyluciferase and light in the presence of ATP and other cofactors, including Mg²⁺ and molecular oxygen. Thus, the assay detects ATP produced by metabolically active viable cells, yielding a luminescent signal that is directly proportional to the total number of cells per well in a 384-well format. The assay was used to screen new nanoemulsion compounds for cell toxicity on murine macrophage (Raw264.7), epithelial cell (TC-1), and dendritic cell (Jaws II) lines. The 50% inhibitory concentration (IC₅₀) for each formulation was calculated after 24 h of NE exposure. The IC₅₀ is defined as the nanoemulsion concentration (percent, wt/wt) yielding 50% inhibition at 24 h for each cell line. For the formulation of NB-201, the IC₅₀ was 0.073% in TC-1 cells.

Garcia A., Fan Y.Y., Vellanki S., Huh E.Y., Vanegas D., Wang S.H, & Lee S.C. (2019). Nanoemulsion as an Effective Treatment against Human-Pathogenic Fungi. mSphere, 4(6), e00729-19.

+ Open protocol

+ Expand

High-Throughput Luciferase Assay Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Recombinant luciferase at 10–15 mg/ml (Promega) was diluted 10⁶ times in complete medium and dispensed, 12 µl/well, in white 384-well plates (Proxiplate, Perkin Elmer) pre-spotted with compounds. After 1 h of incubation, 6 µl of Bright-Glo reagent (Promega) was added to the wells. Following a 3-min incubation, luciferase activity was measured using a Synergy 2 multi-purpose plate reader (Biotek, Winooski, VT). Each plate contained vehicle controls (0.1% DMSO) and wells that contain either no luciferase or no Bright-Glo. The average of these controls was used for the normalization of the compound signals.

Miglianico M., Bolscher J.M., Vos M.W., Koolen K.J., de Bruijni M., Rajagopal D.S., Chen E., Kiczun M., Gray D., Campo B., Sauerwein R.W, & Dechering K.J. (2023). Assessment of the drugability of initial malaria infection through miniaturized sporozoite assays and high-throughput screening. Communications Biology, 6, 216.

+ Open protocol

+ Expand

Splenocyte Proliferation and Cytokine Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mice were euthanized, spleens harvested aseptically, finely minced in 10
ml RPMI, and filtered through a 40 μm cell strainer to remove debris. The
cell suspension was transferred to a 15 ml tube and centrifuged at 200 ×
g for 5 minutes. Cells were resuspended at 5×10⁶ viable
cells/ml in RPMI with 10% FBS and penicillin and streptomycin. Concavalin A
(ConA), a non-specific mitogen, was used as a positive control for the
proliferative ability of splenocytes in the assay. Samples were set up in
triplicate in a 96-well plate with 1 μg recombinant luciferase (Promega)
or ConA (1 μg), 100 μl of growth medium, and 100 μl of
spleen cell suspension. Antibody pairs were used and performed per manufacturer
instructions of analysis of interleuken-2 (IL-2) and interferon-γ
(IFN-γ) (MabTech Inc., Cincinnati, OH). Spots were analyzed by using a
ImmunoSpot/BioSpot UV Analyzer (CTL Analyzers, Shaker Heights, OH). Change as
compared to unstimulated negative control cells was plotted.

Tai D.S., Hu C., Kim E.H, & Lipshutz G.S. (2015). Augmentation of Transgene-Encoded Protein After Neonatal Injection of Adeno-Associated Virus Improves Hepatic Copy Number Without Immune Responses. Pediatric research, 78(3), 239-246.

+ Open protocol

+ Expand

Luciferase and β-Galactosidase Assays

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed by the addition of 100 μl of passive lysis buffer (Promega/Biotium) to each well and shaken for >15min. Luciferase assay was carried out as described in (Chan and Egan, 2005 (link); Chan and Egan, 2009 (link); Egan et al., 2013 (link); Chan et al., 2021 (link)) in a buffer containing 0.0165 M glycylglycine, 0.01 M MgSO₄, 2.65 mM EGTA, 10.5 mM potassium phosphate, 1.4 mM adenosine 5′-triphosphate, 0.86 mM dithiothreitol (DTT), 0.175 mg/ml bovine serum albumin (BSA), and 0.035 mM luciferin (Promega) using 50 μl of the lysate and a luminometer (Berthold Technologies, Germany). A standard curve was constructed using recombinant luciferase (Promega). Beta-galactosidase assay was carried out as described in (Chan and Egan, 2005 (link); Chan and Egan, 2009 (link); Egan et al., 2013 (link)) in a buffer containing 100 mM sodium phosphate pH7.3, 1mM MgCl₂, 50 mM β-mercaptoethanol and 0.665 mg/ml o-nitrophenyl β-D-galactopyranoside (Sigma) using 20 μl of the lysate and read at 420 nm using a plate reader (Bio-Tek Synergy HT). Luciferase activity was normalized against β-galactosidase activity.

, & Chan S.W. (2022). Fusion assays for screening of fusion inhibitors targeting SARS-CoV-2 entry and syncytia formation. Frontiers in Pharmacology, 13, 1007527.

+ Open protocol

+ Expand

ELISA for Antigen-Specific Antibody Titers

Check if the same lab product or an alternative is used in the 5 most similar protocols

High-binding Nunc Maxisorp™ flat bottom 96-well plates were coated with 100 µL/well of 2 µg/mL of recombinant luciferase (Promega) in PBS. After overnight incubation at 4 °C, plates were washed with PBS containing 0.05% Tween 20 (PBS-T) and blocked with 3% (w/v) Bovine Serum Albumin (IgG-Free, Protease-Free) (Jackson Immuno Research, West Grove, PA, USA) in PBS-T. After washing, two-fold serial dilutions of the mouse serum in blocking buffer were added for one hour at 37 °C. Plates were washed again, then HRP-conjugated goat anti-mouse IgG (Cytiva, Marlborough, MA, USA), HRP-conjugated goat anti-mouse IgG1 (Jackson Immuno Research, West Grove, PA, USA), or HRP-conjugated goat anti-mouse IgG2a (Jackson Immuno Research) were added at a 1:2000 dilution and incubated for one hour at 37 °C. 75 µL of tetramethylbenzidine (TMB) substrate (Cell Signaling Technology, Danvers, MA, USA) was added after washing and plates were incubated at room temperature for five minutes. The reaction was terminated by addition of 0.16 M sulfuric acid and absorbance was read at 450 nm on a spectrophotometer. Endpoint antibody titers were expressed as the reciprocals of the final detectable dilution, with a cut-off value defined as the mean of all wells containing serum from mice in the empty particle control group plus three times the standard deviation.

Zhang W., Pfeifle A., Lansdell C., Frahm G., Cecillon J., Tamming L., Gravel C., Gao J., Thulasi Raman S.N., Wang L., Sauve S., Rosu-Myles M., Li X, & Johnston M.J. (2023). The Expression Kinetics and Immunogenicity of Lipid Nanoparticles Delivering Plasmid DNA and mRNA in Mice. Vaccines, 11(10), 1580.

+ Open protocol

+ Expand

Evaluation of Micelle-Mediated Gene Delivery

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human cervical cancer line HeLa cells (ATCC) were maintained in Dulbecco's Modified Eagle's Medium supplemented with 10% fetal bovine serum at 37 °C and 5% CO₂. Cells were seeded in 48-well plates at a density of 2 × 10⁴ cells/well and incubated overnight prior to further use. Micelles with different shapes were added to each well at a dose of 1 μg of plasmid DNA. After 4 h of incubation, the culture media were refreshed. The culture media were removed after 48 h and cells were lysed with 100 μL of reporter lysis buffer for each well (Promega, Madison, WI), and subjected to two freeze–thaw cycles. Twenty μL of cell lysate supernatant was mixed with 100 μL of luciferase substrate (Promega), and the light units were measured on a luminometer (20/20n Single Tube luminometer, Turner BioSystems, Sunnyvale, CA). The luciferase activity was converted to the amount of luciferase using recombinant luciferase (Promega) as the standard, and normalized against protein content using the BCA protein assay (Thermo Scientific, Rockford, IL)

We Z., Ren Y., Williford J.M., Qu W., Huang K., Ng S., Mao H.Q, & Luijten E. (2015). Simulation and Experimental Assembly of DNA–Graft Copolymer Micelles with Controlled Morphology. ACS biomaterials science & engineering, 1(6), 448-455.

+ Open protocol

+ Expand

Splenocyte Proliferation and Cytokine Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

+ Open protocol

+ Expand

AAV-mediated Gene Delivery in Mice

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Procedures were approved by the University of California, Los Angeles
Committee on Animal Research. FVB/N female and male mice were purchased from
Charles River Breeding Laboratories (Wilmington, MA). MyD88 knockout mice and
C57BL/6 controls were from Jackson Laboratories (Bar Harbor, ME). FVB/N mice
were used for all studies otherwise. At birth, an intravenous injection of
3×10¹² AAV genome copies (gc)/kg in 50 μl of normal
saline was performed as previously described (6 (link)). Adult mice received alum/luciferase by intraperitoneal (IP)
injection to the right lower abdomen. AAV was delivered in adults as
3×10¹² gc/kg in 200 μl of normal saline by tail
vein injection.
Recombinant luciferase (Promega, Madison, WI) was mixed with a
pre-formulated aqueous solution of aluminum hydroxide and magnesium hydroxide
(Imject Alum, Pierce, Rockford, IL) in a 1:1 ratio according to the
manufacturer’s instructions. A volume of 200 μl (including 1
μg of Recombinant luciferase) was administered IP to each mouse.

+ Open protocol

+ Expand

Quantifying Promoter Efficiency by Luciferase Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

24 hours after seeding into 6-well plates, cells from each promoter haplotype variant of the reporter gene cell lines were washed with PBS before lysis with Glo lysis buffer (Promega). Lysates were assayed in triplicate for luciferase expression using the Bright-Glo assay kit (Promega), using a known concentration of recombinant luciferase (Promega) as a reference sample to normalise luminescence data. Protein concentration (μg/ml) of lysates was quantified in triplicate using the BCA protein assay kit (Perbio). Promoter efficiency was quantified by comparing specific activity (normalised luminescence/protein concentration of lysates) for the different promoter haplotypes.

Lovewell T.R., McDonagh A.J., Messenger A.G., Azzouz M, & Tazi-Ahnini R. (2015). The AIRE -230Y Polymorphism Affects AIRE Transcriptional Activity: Potential Influence on AIRE Function in the Thymus. PLoS ONE, 10(5), e0127476.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!