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Gwiz luc

Manufactured by Aldevron
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The GWiz-Luc is a laboratory instrument used for luminescence detection. It is designed to measure and quantify luciferase reporter gene expression in biological samples.

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Lab products found in correlation

Gwiz gfp, by Aldevron (4 mentions) Agarose, by Thermo Fisher Scientific (1 mentions) Generuler 1 kb dna ladder, by Thermo Fisher Scientific (1 mentions) Celltiter glo 2.0 reagent, by Promega (1 mentions) Branched polyethylenimine pei, by Merck Group (1 mentions) Ethidium bromide 1 solution, by Thermo Fisher Scientific (1 mentions) Dna loading dye, by Thermo Fisher Scientific (1 mentions) Cell culture lysis 5x reagent, by Promega (1 mentions) Adenosine triphosphate (atp), by MP Biomedicals (1 mentions) Heparin sodium salt, by MP Biomedicals (1 mentions) Ebioscience 7 aad viability staining solution, by Thermo Fisher Scientific (1 mentions) Annexin 5 alexa fluor 647 conjugate, by Thermo Fisher Scientific (1 mentions) Pcmv6 xl4 vector, by OriGene (1 mentions) D luciferin, by GoldBio (1 mentions) Limulus amebocyte lysate assay, by Aldevron (1 mentions) Heparin, by Merck Group (1 mentions) Sodium nitrite, by Merck Group (1 mentions) 3 5 dinitrosalicylic acid, by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) 5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (1 mentions)

Spelling variants of this product

GWiz™ Luc plasmid (3 citations)

8 protocols using gwiz luc

1

Plasmid-mediated gene delivery protocol

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gWiz plasmid containing luciferase (gWiz-Luc), or Green Fluorescence Protein reporter gene (gWiz-GFP) were purchased from Aldevron (Fargo, ND) as 5 mg/ml aqueous solution and used without further purification. Plasmid DNA encoding the human interferon-beta1 (IFN-β1) inserted into a pCMV6-XL4 vector was purchased from OriGene Technologies Inc. (Rockville, MD) and used without further purification. Polyethylenimine branched (PEI, molecular weight 25,000 Da) was purchased from Sigma-Aldrich (St. Louis, MO). Ethidium Bromide: 1% solution, and agarose were procured from Fisher Bioreagents (Fair Lawn, NJ). GeneRuler 1 kb (DNA ladder) and 6x DNA loading dye were purchased from Thermo Scientific (Vilnius, Lithuania). Cell Titer Glo 2.0 reagent and cell culture lysis 5x reagent were from Promega (Madison, WI). ATP, Lucifer Yellow CH dilithium, and heparin sodium salt were purchased from MP Biomedicals (Illkirch, France). Pierce BCA protein assay kit for total protein measurement was purchased from Thermo Scientific (Rockford, IL). Annexin V Alexa Fluor 647 conjugate and e-Bioscience 7-AAD viability staining solution were procured from Thermo Fisher Scientific (Carlsbad, CA). All the chemicals used for the experiments were obtained as molecular biology grade, DNase, RNase, and protease-free materials.
Dave K.M., Han L., Jackson M.A., Kadlecik L., Duvall C.L, & Manickam D.S. (2020). DNA Polyplexes of a Phosphorylcholine-Based Zwitterionic Polymer for Gene Delivery. Pharmaceutical research, 37(9), 176.
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2

Molecular Tools for Cell Analysis

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The reporter plasmids encoded luciferase (gWiz-Luc) and green fluorescent protein (gWiz-GFP) were purchased from Aldevron (Fargo, ND). CPA, a reagent for intracellular calcium depletion, was obtained from Iurii Semennov (Old Dominion University). D-Luciferin, which was utilized for luciferase assay or live cell imaging, was purchased from Goldbio Technology (St Louis, MO). WST-1 for cell viability assay was obtained from Roche Applied Science (Indianapolis, IN).
Guo S., Jackson D.L., Burcus N.I., Chen Y.J., Xiao S, & Heller R. (2014). Gene electrotransfer enhanced by nanosecond pulsed electric fields. Molecular Therapy. Methods & Clinical Development, 1, 14043-.
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3

Plasmid DNA for Luciferase and VEGF-A

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Plasmid DNA encoding luciferase, gWizLuc, was purchased from Aldevron (Fargo, ND). Plasmid DNA encoding human VEGF-A165 tagged with a DDK tag (pVEGF-A-DDK) purchased from OriGene (Rockville, MD). Plasmid DNA was suspended in sterile saline at 2 mg/mL by Aldevron. Endotoxin levels were < 0.1 EU/μg plasmid, confirmed by Aldevron via a Limulus Amebocyte Lysate assay.
Bulysheva A., Hornef J., Edelblute C., Jiang C., Schoenbach K., Lundberg C., Malik M.A, & Heller R. (2018). Coalesced thermal and electrotransfer mediated delivery of plasmid DNA to the skin. Bioelectrochemistry (Amsterdam, Netherlands), 125, 127-133.
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4

Lipid-based Nanoparticle Formulation and Characterization

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Lipids including 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-mPEG(2 kDa)), 1,2-distearoyl-sn-glycero-3-phosphoethanol-amine-N-[maleimide(polyethylene glycol)-2000] (ammonium salt) (DSPE-PEG(2 kDa)-MAL) and 1,2-dioleoyl-3-trimethy-lammonium-propane (DOTAP) were obtained from Avanti Polar Lipids and used without further purification. Heterobifunctional cross-linker 3-(2-pyridyldithio)propionyl hydrazide) (PDPH) was obtained from Pierce. Heparin, sodium nitrite, 3,5-dinitrosalicylic acid, tris(bipyridine)ruthenium(II) chloride), 5,5′-dithiobis(2-nitrobenzoic acid) (Ellman's reagent), tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl), and Triton X-100 were obtained from Sigma-Aldrich and used without further purification. Plasmid DNA encoding for reporter proteins firefly luciferase (gWiz-Luc) and GFP (gWiz-GFP) were obtained from Aldevron.
Chertok B., Langer R, & Anderson D.G. (2016). Spatial Control of Gene Expression by Nanocarriers Using Heparin Masking and Ultrasound-Targeted Microbubble Destruction. ACS nano, 10(8), 7267-7278.
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5

HaCaT Cell Transfection with Plasmids

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The human Keratinocyte cell line, HaCaT, was used for all in vitro experimental studies. Cells were cultured in DMEM + pen-Strep and 10% FBS. Cells were harvested from flasks with 0.25% Trypsin-EDTA and resuspended at 5 million cells/ml in complete media.
Plasmids: 5 μl of either Gwizluc or GwizGFP (Aldevron) at 2 mg/ml were used in this study to evaluate gene expression.
Donate A., Burcus N., Schoenbach K, & Heller R. (2014). Application of increased temperature from an exogenous source to enhance gene electrotransfer. Bioelectrochemistry (Amsterdam, Netherlands), 103, 120-123.
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6

Cellular Uptake Mechanisms Evaluation

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1,4-Butanediol diacrylate and 5-amino-1-pentanol were purchased from Alfa Aesar (Ward Hill, MA, USA). Dodecylamine was purchased from Sigma-Aldrich (St. Louis, MO, USA). (PEO)4-bisamine (“122”) was acquired from Molecular Biosciences (Boulder, CO, USA). All chemical reagents were used without further purification. Plasmids encoding green fluorescent protein (gWiz-GFP) and firefly luciferase (gWiz-Luc) were purchased from Aldevron (Fargo, ND, USA). jetPEI (Polyplus Transfection, Illkirch, France) and Lipofectamine 2000 were purchased from VWR (Radnor, PA, USA) and Invitrogen (Carlsbad, CA, USA), respectively. Transferrin, cholera toxin B, and 10 000 MW dextran, each labeled with AlexaFluor 647, were purchased from Invitrogen. Cytochalasin D, dynasore hydrate, chlorpromazine hydrochloride, filipin III, genistein, methyl-β-cyclodextrin, 5-(N-ethyl-N-isopropyl)amiloride, and U18666A were obtained from Sigma-Aldrich. Immortalized mouse embryonic fibroblast cell lines were cultured in DMEM (Invitrogen) supplemented with 10% fetal bovine serum (Invitrogen).
Eltoukhy A.A., Sahay G., Cunningham J.M, & Anderson D.G. (2014). Niemann-Pick C1 Affects the Gene Delivery Efficacy of Degradable Polymeric Nanoparticles. ACS Nano, 8(8), 7905-7913.
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7

Synthesis and Characterization of Cationic Polymers

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1,1′-Carbonyldiimidazole
(CDI), bis(2-hydroxyethyl) disulfide (BHED), spermidine (SPM) trihydrochloride,
putrescine (PUT) dihydrochloride, and branched polyethylenimine (PEI,
25 kDa) were purchased from Sigma-Aldrich (St. Louis, MO). Dichloromethane
(DCM) (99.9%, extra dry, AcroSeal), tetrahydrofuran (THF, 99.85%,
extra dry, AcroSeal), 5-sulfosalicylic acid dihydrate (SSA), 1,7-diaminoheptane
(DAH), dansyl chloride (5-dimethylamino-1-naphthalenesulfonyl chloride,
98%), and 1,6-hexanediol were from Acros Organics (Fair Lawn, NJ). l-Proline was from Alfa Aesar (Ward Hill, MA). Spermine (SPM)
was from MP Biomedicals (Santa Ana, CA). BENSpm was synthesized by
following a previously described procedure.22 (link) Plasmid DNA containing a luciferase reporter gene (gWiz-Luc) was
from Aldevron (Fargo, ND), and human tumor necrosis factor (TNFα)
plasmid DNA was obtained from InvivoGen (San Diego, CA). Dulbecco’s
modified Eagle’s medium (DMEM), Eagle’s minimum essential
medium (EMEM), Dulbecco’s phosphate-buffered saline (PBS),
fetal bovine serum (FBS), l-glutamine, and penicillin–streptomycin
were from Thermo Scientific (Waltham, MA). RT-PCR primers were purchased
from Invitrogen (Carlsbad, CA). All other reagents and chemicals were
obtained from Fisher Scientific or VWR International unless otherwise
stated.
Zhu Y., Li J., Kanvinde S., Lin Z., Hazeldine S., Singh R.K, & Oupický D. (2014). Self-Immolative Polycations as Gene Delivery Vectors and Prodrugs Targeting Polyamine Metabolism in Cancer. Molecular Pharmaceutics, 12(2), 332-341.
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8

Plasmid Light Scattering Analysis

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A 1.5 μM solution of plasmid gWiz-Luc (6732 bp; Aldevron, Fargo, ND, USA) was prepared in 10 mM cacodylate buffer (pH 7.2) in a total volume of 2.5 ml. Small volumes (<10 μl) of metallohelices were added to the DNA solution in a 1 cm quartz cuvette to obtain the desired concentration. The mixture was thoroughly mixed by pipetting and left undisturbed for 3 min at RT. Total intensity light scattering was measured at 90° angle with respect to the incident beam by using Varian Cary Eclipse spectrofluorophotometer with the following parameters: the excitation and emission wavelengths were set to 305 nm, the excitation and emission slit widths were 5 nm, and the averaging time was set to 3 s.
Malina J., Kostrhunova H., Novohradsky V., Scott P, & Brabec V. (2022). Metallohelix vectors for efficient gene delivery via cationic DNA nanoparticles. Nucleic Acids Research, 50(2), 674-683.
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