PML is required for p73 transcriptional and proapoptotic activity. (A) A bax promoter-driven luciferase reporter plasmid (bax-Luc) alone or in combination with
There was a positive correlation between CEA levels and CA 15-3 levels and patient prognosis. A high specificity for B-cells was confirmed by assaying for luciferase reporter gene expression from a plasmid containing an analogous combination of immunoglobulin heavy chain regulatory ...
Luciferase assays: Use a secreted luciferase reporter for promoter studies in bacteria, cultured cells, and transgenic plants or animals.
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
Both the luciferase gene and a human housekeeping gene ACTB (both expressed only in human cells) were present only in the injected kidney tissue, as confirmed by the absence of the both housekeeping ACTB and luciferase gene in non-injected kidneys (Figure 3D ...
Thermo Scientific™ Gaussia Luciferase Reporter Assay Vectors pMCS-Gaussia Luc Vector Thermo Scientific™ Gaussia Luciferase Reporter Assay Vectors...
Thermo Scientific™ Pierce™ Gaussia Luciferase Flash Assay Kit 100-rxn kit Thermo Scientific™ Pierce™ Gaussia Luciferase Flash Assay...
Gaussia Luciferase (GLuc) reporter gene offers bright bioluminescence, either as a standalone expression monitor or as a fusion partner with other protein.
The half-life of GLuc remains unclear. All GLuc-expressing vectors available for mammalian expression at NEB harbor the humanized codons for GLuc in which the secreted GLuc has activity similar to that of the native protein. Once GLuc is secreted into the culture medium, it is very stable. For example, ~20% of the GLuc activity was detectable after incubating a GLuc-containing sample at 99˚C for 15 minutes ...
The original aim of this thesis was to utilise Vibrio harveyi luciferase as a reporter of the expression of cell division genes during the cell cycle. Several plasmids expressing luxAB genes from ftsZ promoters were constructed. To achieve maximal luciferase expression, the ribosome binding site in front of the luxA gene was improved, which led to increased expression of luciferase. The level of expression of the improved luciferase reporter from plasmids was sufficiently high to be detected in single cells, although not high enough to be used in lower copy number constructs. However, luciferase activity showed significant fluctuations that did not appear to be linked to cell cycle events. These fluctuations made the detection of any cell cycle related changes in luciferase expression impossible. Another direction of this thesis is represented by the studies on the topology of the cell shape determining RodA protein. The ampicillin resistance levels were measured in 52 fusions with the topology ...
The pGL3 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These factors may be cis-acting, such as promoters and enhancers, or trans-acting, such as various DNA-binding factors. The backbone of the pGL2 Luciferase Reporter Vectors was redesigned for the pGL3 Vectors for increased expression, and contains a modified coding region for firefly (Photinus pyralis) luciferase that has been optimized for monitoring transcriptional activity in transfected eukaryotic cells. The assay of this genetic reporter is rapid, sensitive and quantitative. In addition, the Luciferase Reporter Vectors contain numerous features aiding in the structural characterization of the putative regulatory sequences under investigation.. ...
To confirm the functionality of LODER-driven siRNA in vivo, we tested the ability of a LODER containing siRNA targeting the luciferase gene, siLuc LODER, to reduce luciferase expression in normal and tumor tissues constitutively expressing the luciferase gene. We therefore implanted empty or siLuc LODERs into the livers of transgenic mice expressing the luciferase gene (MUP-Luc) in liver cells (19). In vivo imaging results showed that siLuc LODERs led to a significant decrease in luciferase levels compared with empty LODERs (Fig. S5A). Next, LODERs carrying siLuc or siGFP were implanted into CT26 cell-derived s.c. synograft tumors. Three days after implantation, measurements of luciferase activity in vivo revealed that siLuc released from the LODERs inhibited luciferase expression (Fig. S5B). This decrease in luciferase activity was not correlated to nonspecific effects on tumor growth, as tumor weights were similar within both groups (Fig. S5C). Together these results show that siRNA is ...
Gentaur molecular products has all kinds of products like :search , Panomics \ AR Luciferase Reporter Vector \ LR0007 for more molecular products just contact us
Gentaur molecular products has all kinds of products like :search , Signosis 2011 \ TF Luciferase Reporter Vectors \ LR-2XXX for more molecular products just contact us
Material and Methods: miR-150 expression was quantified by qRT-PCR in two MIBC cell lines (5637 and T24). After successful miR-150 inhibition by transfection, MTS and transwell assays were used to assess the MIBCs cisplatin sensitivity and cell invasiveness, respectively. The TargetScan database and a luciferase reporter system were used to identify whether the programmed cell death 4 protein (PDCD4) is a direct target of miR-150 in MIBC cells ...
The LightSwitch 3´UTR Reporter Collection from Active Motif include over 12,000 3´UTRs available as transfection-ready luciferase reporter vectors for use in reporter assays.
IL-8 is a direct target of miR-106a. (a) Up: potential sites in IL-8 3′UTR targeted by miR-106a. 3′UTR of IL-8 was cloned into a luciferase reporter vector.
Plasmid pGL3-NFAT luciferase from Dr. Jerry Crabtrees lab contains the insert 3x NFAT binding sequence and is published in Nature. 1992 Jun 25. 357(6380):695-7. This plasmid is available through Addgene.
Buy Luciferase recombinant protein-BAF48390 (MBS203675) product datasheet at MyBioSource, Recombinant Proteins. Application: SDS-PAGE
BioAssay record AID 95279 submitted by ChEMBL: Cellular activity was measured by an IL-2 luciferase reporter gene assay on a Jurkat human T cell line activated by anti-CD28 and anti-TCR antibodies.
Dual luciferase assay - proper controls? - posted in Cell Biology: Hey everyone, I am trying to perform a dual luciferase reporter assay in a cell line with promoters established from another cell line. cDNA and protein of the gene are present in both cell lines. Problem: In the new cell line, the luciferase activity for my promoters of interest is below pGL3, whereas TKpGL3 is as highly active as in the other cell line. Renilla activity was equal in all probes. So I am looking for appr...
Bioluminescent Reporter plasmid to express bacterial luciferase (Lux) in Mycobacteria Background. Bioluminescence, the production of light by luciferase-catalysed reactions, is a versatile reporter technology with multiple applications both in vitro and in vivo. In vivo bioluminescence imaging (BLI) represents one of its most outstanding uses by allowing the non-invasive localisation of luciferase-expressing cells within an animal. Applied to the study of infectious diseases, BLI permits the detection of microorganisms from within living animals thus allowing the spatiotemporal study of infection in real-time in the same host. Moreover, using luciferase as a reporter of gene expression, it is possible to establish when and where a gene function is needed, shedding light on bacterial pathogenesis. Finally, BLI constitutes an easy and rapid method to test novel antimicrobial compounds in vivo.. Three main luciferin-luciferase systems have been utilised for BLI. The first system is represented by ...
0140] The in vitro transfection efficiency of complexes prepared with modified bPEI-2 polymer and luciferase reporter gene was investigated using HepG2 and SK-OV-3 cell lines. HepG2 cells were seeded onto 24-well plates at a density of 8×104 cells per 500 microliters per well for luciferase gene delivery. SK-OV-3 cells were seeded onto 24 well plates at a density of 8×104 cells per 500 microliters per well for luciferase gene delivery. After 24 hours, the plating media were replaced with fresh growth media, followed by the drop-wise addition of 50 microliters of complex solution containing 2.5 micrograms luciferase plasmid DNA at various N/P ratios. Following 4 hours of incubation, free complexes were removed by replacing the medium in each well. After a further 68 hours of incubation, the cell culture medium in each well was removed and the cells rinsed once with 0.5 mL of phosphate-buffered saline (PBS, pH 7.4). For luciferase expression assay, 0.2 mL of reporter lysis buffer was added to ...
pGL4.26[luc2/minP/Hygro], pGL4.27[luc2P/minP/Hygro] and pGL4.28[luc2CP/minP/Hygro] Vectors are optimized for expression in mammalian cells. Use these vectors to clone a response element of interest upstream of the minimal promoter and firefly luciferase gene with and without destabilization sequences. Offer hygromycin selection for stable transfection.
The temporal effects of luciferase reaction luminescence have only been discussed in the context of light intensity (flash vs. glow). However, alterations in the color of the light emitted over the course of the luciferase reaction have not been reported. Here, we show a temporal change in the light color emitted during the reaction catalyzed by unmodified firefly luciferase when concentrations of one of the substrates, adenosine triphosphate (ATP), are gradually increased. The temporal color change from green to red occurs within the first few minutes of the luciferase reaction when an ATP-containing solution is either added or synthesized in situ with the aid of an autocatalytic reaction occurring simultaneously ...
Alibaba.com offers 155 luminometer products. About 27% of these are testing equipment, 25% are other measuring & analysing instruments, and 6% are other optics instruments. A wide variety of luminometer options are available to you,
Title:Effects of APC De-Targeting and GAr Modification on the Duration of Luciferase Expression from Plasmid DNA Delivered to Skeletal Muscle. VOLUME: 15 ISSUE: 1. Author(s):Maria C. Subang, Rewas Fatah, Ying Wu, Drew Hannaman, Jason Rice, Claire F. Evans, Yuti Chernajovsky and David Gould. Affiliation:Bone & Joint Research Unit, Queen Mary University of London, William Harvey Research Institute, Charterhouse Square, London EC1M 6BQ, UK.. Keywords:Gene therapy, luciferase, microRNA, plasmid DNA, skeletal muscle, tissue-specific promoter, transgene immunogenicity.. Abstract:Immune responses to expressed foreign transgenes continue to hamper progress of gene therapy development. Translated foreign proteins with intracellular location are generally less accessible to the immune system, nevertheless they can be presented to the immune system through both MHC Class I and Class II pathways. When the foreign protein luciferase was expressed following intramuscular delivery of plasmid DNA in outbred ...
a,b, Western blotting analysis of miR-20a target genes in response to increasing doses of miR-20a mimic (a) or antagomir (b). c,d, RT-qPCR analysis of miR-20a levels in HeLa cells transfected with miR-20a mimic (c) or antagomir (d). e,f, RT-qPCR analysis of DAPK3 mRNA levels in HeLa cells transfected with miR-20a mimic (e) or antagomir (f). g, Luciferase reporter constructs containing the miR-20a MRE from DAPK3 (native) in Renilla 5 UTR or mutated to restore base-pairing in the 5 end (seed) coupled with progressive mismatches (3MM+seed) in the 3 end. h,i, Results of the luciferase reporter assays (h) and quantification of luciferase mRNA (i). j,k, mRNA levels of luciferase reporters containing individual MREs from 4 indicated genes that function in both CDS and 3UTR (j) or those that function only in CDS (k), as shown in main Fig. 2e,f. l,m, Luciferase activities from the reporters containing individual MREs from 4 indicated genes that function only in CDS in response to co-transfected ...
Cells transfected with 8 pmol of luc2-encoding IVT RNA were harvested at different time points to measure luminescence from 1 × 104 cells (Bright-Glo Luciferase Assay Kit for 96-well plates (Promega)). Results are the mean ± SD luminescence. Percent luminescence is relative to the highest luminescence signal obtained in each experiment. (c) Viability, reporter gene expression of iDCs and mDCs after eGFP and luc2 electroporation and phenotype after electroporation are depicted in descending order, respectively. iDCs (left panel) and mDCs (right panel) of 2 different donors were transfected with 10 μg eGFP- or luc2-encoding IVT RNA. Negative controls: cells electroporated without RNA (mock) and unelectroporated (no ep) cells. Cells were harvested at different time points. Viability and HLA-DR, CD83, and eGFP expression levels were determined by flow cytometry. Luciferase activity of 1 × 104 viable cells was measured by luminescence in triplicate ...
Label License, Gaussia luciferase, material transfer license, US Patent, 6232107, 6436682, 6780974, 7045599, 7109315, 7238497, HTS, high throughput screening
The recombinant cell line used in this assay (H1L6.1c2) was generated by stably transfecting the plasmid pGudLuc6.1 into mouse hepatoma (Hepa1c1c7) cells. The pGudLuc6.1 plasmid contains the CYP1A1 dioxin-responsive domain (inclusive of four DREs) upstream of the firefly luciferase gene ...
NanoLuc luciferase brings new capabilities to research applications such as BRET, Reporter Assays, protein:protein interactions, protein:ligand interactions, protein dynamics, genetically-encoded biosensors and in vivo imaging. The small size and extreme brightness of NanoLuc luciferase make it useful in many situations where other luciferases may fail.
A paper published on October 2 in the Journal of Virology describes an exciting development in the world of influenza research-the construction of a luciferase reporter virus that does not affect virulence and can be used to track development and spread of infection in mice.
When I joined the lab team in May I was familiar with lab equipment and testing but I wasnt familiar with luminometers and ATP. I had heard of ATP in my
Our comprehensive collection contains more than 12,000 endogenous human 3′UTR reporter GoClone® constructs that are transfection-ready with no DNA preparation required. Our reporters contain a novel luciferase gene for industry-leading sensitivity ...
Our comprehensive collection contains more than 12,000 endogenous human 3′UTR reporter GoClone® constructs that are transfection-ready with no DNA preparation required. Our reporters contain a novel luciferase gene for industry-leading sensitivity ...
We developed a Gaussia luciferase (Gluc) reporter replicon of West Nile virus (WNV) and used it to quantify viral translation and RNA replication. The advantage of the Gluc replicon is that Gaussia luciferase is secreted into the culture medium from cells transfected with Gluc replicon RNA, and the medium can be assayed directly for luciferase activity. Using a known Flavivirus inhibitor (NITD008), we demonstrated that the Gluc-WNV replicon could be used for antiviral screening. The Gluc-WNV-Rep will be useful for research in antiviral drug development programs, as well as for studying viral replication and pathogenesis of WNV.
Candida albicans is a major fungal pathogen causing life-threatening diseases in immuno-compromised patients. The efficacy of current drugs to combat C. albicans infections is limited, as these infections have a 40-60% mortality rate. There is a real need for novel therapeutic approaches, but such advances require a detailed knowledge of C. albicans and its in vivo pathogenesis. Additionally, any novel antifungal drugs against C. albicans infections will need to be tested for their in vivo efficacy over time. Fungal pathogenesis and drug-mediated resolution studies can both be evaluated using non-invasive in vivo imaging technologies. In the work presented here, we used a codon-optimized firefly luciferase reporter system for detecting C. albicans in mice. We adapted the firefly luciferase in order to improve its maximum emission intensity in the red light range (600-700 nm) as well as to improve its thermostability in mice. All non-invasive in vivo imaging of experimental animals was performed with a
3781 In this study, we examined transfection efficiency of naked plasmid DNA using microbubbles (Optison) with ultrasound in vitro and in vivo experiments as the feasibility of a novel nonviral vector system that transfer naked plasmid DNA into a melanoma. First, we tested the feasibility of ultrasound-mediated transfection of naked plasmid DNA in mouse melanoma B16F1 cell lines. Luciferase plasmid mixed with or without Optison was transfected into cultured mouse melanoma cells using ultrasound (1MHz; 0.4 W2) for 1 sec. Interestingly, luciferase activity was dramatically increased in cells treated with Optison, while few luciferase activity could be detected without Optison (P,0.05). We then transfected luciferase plasmid mixed with Optison by means of therapeutic ultrasound into mouse melanoma tumor. Twenty four hours after transfection, luciferase activity, transfected with Optison and ultrasound, was also higher than that of plasmid alone. Finally, we examined the feasibility of therapeutic ...
The bacterial luciferase gene cassette (lux) is unique among bioluminescent bioreporter systems due to its ability to synthesize and/or scavenge all of the substrate compounds required for its production of light. As a result, the lux system has the unique ability to autonomously produce a luminescent signal, either continuously or in response to the presence of a specific trigger, across a wide array of organismal hosts. While originally employed extensively as a bacterial bioreporter system for the detection of specific chemical signals in environmental samples, the use of lux as a bioreporter technology has continuously expanded over the last 30 years to include expression in eukaryotic cells such as Saccharomyces cerevisiae and even human cell lines as well. Under these conditions, the lux system has been developed for use as a biomedical detection tool for toxicity screening and visualization of tumors in small animal models. As the technologies for lux signal detection continue to improve, it is
FIG 12 HRV16 infection does not block secretion of the Gaussia luciferase reporter. HeLa Ohio cells stably expressing the naturally secreted Gaussia luciferase (HeLa-Gluc) were infected with HRV16 (red squares) at an MOI of 20 for 1 h, followed by culture for the indicated periods of time up to 7 h postinfection (Time hpi). As controls, HeLa-Gluc cells were uninfected (blue circles) or uninfected but treated with BFA (green triangles). The cell culture medium was removed and replaced every 30 min, and the cells were harvested at 1-h intervals. The Gluc activity then was measured in the culture media and cell lysates. The cumulative secretion over time is calculated by adding each successive 30-min culture media sample to the previous total. Panel A shows Gluc activity in the cell lysate, and panel B shows secreted Gluc activity in the cell culture medium. Each point represents the mean relative light units (RLU) (± standard deviations) as a measure of Gluc activity from triplicate assay points ...
Figure 5 Transcriptional and DNA binding effects of BCL11A homomeric and BCL11A-BCL6 heteromeric complexes on artificial target genes. (A) Gal4DBD-BCL11A isoforms repress Gal4-mediated Firefly luciferase activity of pG5luc in BCL1 and M12.4 B cells or in HEK293 cells. Dual luciferase assays were performed 48 hr post-transfection into the indicated cell lines on whole cell lysates shown by Western blotting to contain approximately equal amounts of Gal4-BCL11A isoforms (data not shown). Values are the average of three independent experiments and are expressed as percent of those obtained using the reporter construct and Gal4DBD alone after normalization of transfection efficiency for the activity of Renilla luciferase. (B) Repression by BCL6, but not BCL11A, is TSA sensitive. pG5luc and Gal4DBD-BCL11A isoform fusions or FLAG-BCL6 along with a firefly luciferase reporter vector containing 5×-BCL6 binding sites upstream of the minimal SV40 promoter (5XBCL6-SV40-Luc; REF), and a Renilla luciferase ...
Continuous increase in the number and the variety of anthropogenic sources of electromagnetic radiation causes a high interest in studying the effects ultrahigh frequency on living organisms. In the present research inflence of UHF EMR (15 W, 2.45 GHz) for 5 and 15 min on morphological and genetic peculiarities of Photobacterium phosphoreum colonies was studied. It has been revealed that UHF EMR affected colony growth parameters, induced transcriptional activity of luciferase encoding gene expression and that the effect was depended on exposure duration. The subsequent cultivation of bacteria during a two week period after treatment showed maintaining of the increased luxb mRNA level in irradiated colonies. Opposite bacterial stress responses were detected to UHF EMR and elevated temperature treatments that assumed UHF EMR comprised of not only thermal but specifi component of non-thermal nature.. The Opened International University of Human Development Ukraine ...
The attractiveness of secreted luciferases as reporters is a strong stimulus for the investigation and exploitation of new bioluminescent systems. Metridia longa is a small luminous marine copepod (Fig. 3D). The bioluminescence originates as a secretion from epidermal glands located in the head part and abdomen in response to mechanical, electrical, or chemical stimuli. Bioluminescence in Metridia longa may well serve as a defense mechanism against predators; the release of a luminous bolus from the animal is accompanied by rapid swimming that displaces the copepod away from its "glowing phantom". This luciferase emits light at a peak of 480 nm with a broad emission spectrum extending to 600 nm. Gaussia luciferase has been cloned, overexpressed in bacteria, and used as a sensitive analytical reporter for hybridization assays and monitoring of cellular expression in culture and in vivo (Tannous et al. 2005). 7 Coelenterazine Dependent Luciferases 9 Fig. 3 Origins of Glow-light reporter genes used ...
The pCMV-GLuc 2 Control Plasmid is a mammalian expression vector that encodes the secreted luciferase from the copepod Gaussia princeps as a reporter, under the control of the constitutive CMV (cytomegalovirus) promoter. Gaussia luciferase (GLuc) is a 19 kDa protein encoded by a humanized sequence, and it contains a native signal peptide at the N-terminus that allows it to be secreted from mammalian cells into the cell culture medium. A neomycin resistance gene under the control of an SV40 promoter allows selection for stable integration of the plasmid into the mammalian cell genome using G418.
Since GARP appears to regulate TGF-1 secretion by T cells and TGF-one is essential for the suppressive perform of Treg cells, we sought to identify mechanisms
D-Luciferin Firefly, free acid 4,5-Dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid Luciferin is a common bioluminescent reporter used for in-vivo imaging of the expression of the luc marker gene . It is the substrate for the Firefly luciferase enzyme which utilizes ATP a ...
Andrew C. Liu is the author of this article in the Journal of Visualized Experiments: Monitoring Cell-autonomous Circadian Clock Rhythms of Gene Expression Using Luciferase Bioluminescence Reporters
We have studied the PAC-2 cell line extensively 6,7,14,15 and so have determined a set of electroporation conditions that provide optimal transfec-tion
To further validate direct binding and targeting by miR-10b, we constructed Renilla luciferase reporters containing either wild type or mutated 3′-UTRs of these 4 target genes. Mutations were designed within the miR-10b seed-binding regions of the 3′-UTRs, as indicated in Supplementary Fig. S5, to disrupt the predicted binding. Reporter activities were quantified and normalized to nontargeted Firefly luciferase activity. All 4 reporters containing the wild-type 3′-UTRs showed notable derepression in response to inhibition of miR-10b (Fig. 4C), indicating that miR-10b regulates those 4 genes via their 3′-UTRs. Mutations within the miR-10b binding sites of Bim, TFAP2C, p21, and p16 3′-UTRs partially abolished the responsiveness of the corresponding reporters to the miR-10b inhibitor. These results indicate that these sites are indeed critical for miR-10b binding and mediate its regulation.. Additional evidence supporting regulation of Bim, TFAP2C, and p21 by miR-10b in GBM was obtained ...