Search results for: DNM-9602A Microplate reader
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Highly elastomeric photocurable silk hydrogels.A photocurable silk fibroin hydrogel is prepared, for the first time, using natural silk protein fibroin and biophotosensitizer riboflavin. Riboflavin is excited by ultraviolet light to generate a triplet state which is transferred to produce active oxygen radicals with singlet oxygen as the main component. Active oxygen radicals can induce chemical cross-linking of amino-, phenol- and other groups in the silk fibroin macromolecules to form a photocurable hydrogel. The different biophysical characterizations of the gelation of this modified fibroin protein solution were studied by using Fourier transform infrared spectroscopy, X-ray diffraction, scanning electron microscopy, microplate reader and texture analyzer. The aggregate structures, surface morphologies, mechanical properties, light transmission and degradation properties of the gel were studied. The investigations showed that the silk fibroin/riboflavin hydrogels predominantly have random coils or alpha helix structures. These gels show resilience up to 90% after 80% compression and a light transmission of up to 97%. The cell culture experiment exhibits that the hydrogel has a satisfactory cytocompatibility.
Beta Amyloid (1 42) High StayBrite™ Highly Stabl Apoptosis antibody array Cell cycle antibody array Cytokine antibody array i Master antibody array is Signal transduction antib AKT Phospho-Specific Arra AKT PKB Signaling Phospho AMPK Signaling Phospho-Sp Apoptosis Phospho-Specifi Cancer Apoptosis Phospho-
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A strongly fluorescing anaerobic reporter and protein-tagging system for organisms based on the Fluorescence-Activating and Absorption-Shifting Tag (FAST) protein.Visualizing protein localization and characterizing gene-expression activity in live cells is limited for lack of a real-time, highly-fluorescent, oxygen-independent reporter systems. Enzymatic reporter systems have been used successfully for many years in however, these assays do not allow for real-time analysis of gene-expression activity with flow cytometry or for visualizing protein localization through fusion proteins. Commonly used fluorescent reporter proteins require oxygen for chromophore maturation, and cannot be used for most strictly anaerobic organisms. Here we show that the fluorescence-activating protein (FAST), when associated with the fluorogenic ligand 4-hydroxy-3-methylbenzylidene-rhodanine (HMBR, now commercially available) and other commercially available ligands, is highly fluorescent in under anaerobic conditions. Using flow cytometry and a fluorescent microplate reader, we demonstrate FAST as a reporter system by employing the promoters of the thiolase (), acetoacetate decarboxylase () and phosphotransbutyrylase () metabolic genes, as well as a mutant P and modified RBS versions of P and P Flow-cytometry based sorting was efficient and fast in sorting FAST expressing cells, and positively and negatively sorted cells could be effectively re-cultured. FAST was also used to tag and examine protein localization of the predicted cell-division FtsZ-partner protein, ZapA, to visualize the divisome localization in live cells. Our findings suggest that FAST can be used to further investigate divisomes and more broadly the localization and expression levels of other proteins in organisms, thus enabling to carry out cell-biology studies in these organisms. FAST and the fluorogenic ligand HMBR is characterized as a successful, highly-fluorescent reporter system in FAST can be used to distinguish between promoters in live cells using flow cytometry or a fluorescent microplate reader, and can be used to tag and examine protein localization in live, anaerobically grown cells. Given that FAST is highly fluorescent under anaerobic conditions, it can be used in several applications of this and likely many organisms and other strict anaerobes including studies involving cell sorting, sporulation dynamics, and population characterization in pure as well as mixed cultures such as those in various native or synthetic microbiomes and syntrophic cultures.
1990 related Products with: A strongly fluorescing anaerobic reporter and protein-tagging system for organisms based on the Fluorescence-Activating and Absorption-Shifting Tag (FAST) protein.Rabbit Anti-Rat Androgen Bone Morphogenetic Protei Amplite™ Fluorimetric F Green Fluorescence Protei NATIVE HUMAN PROLACTIN, P Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy Rabbit Anti-FLAP 5-lipoxy
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Standardization of a resazurin-based assay for the evaluation of metabolic activity in oral squamous carcinoma and glioblastoma cells.The widely accepted resazurin-based assay can be used, priory to in vivo studies, as an inexpensive method to determine cytotoxicity. The aim of this study was to evaluate and standardize the assay conditions for oral squamous cancer cell (OSCC) and glioblastoma (U87-MG) lines by UV-visible spectroscopy. The cells were treated with 25 µgmL of resazurin sodium salt and then incubated for 4 hours, 6 hours, and 6.5 hours. All absorbance measurements were carried out at 21 ± 1 °C on a spectrophotometer with a microplate reader. After 4 -hs of incubation, resazurin was completely reduced by OSCC cells, as demonstrated by the suppression of the absorbance at 380 nm. However, the U87-MG cells needed 6.5 hours of incubation to demonstrate the same behavior. The Statistical analysis did not indicate significant differences between the OSCC and U87-MG cell lines' viability after 4 and 6.5 hours respectively. We concluded that spectroscopic analysis is an efficient method for the standardization of the resazurin assay. In addition, without the implementation of suitable protocols, there could be an increase in the chance of errors or false positives or negatives that would reduce the usefulness of the data.
2101 related Products with: Standardization of a resazurin-based assay for the evaluation of metabolic activity in oral squamous carcinoma and glioblastoma cells.MarkerGeneTM Live Dead As MarkerGene™ LysoLive™ Cell Meter™ Fluorimetri Cell Meter™ Fluorimetri Alkaline Phospatase (ALP) GLP 1 ELISA Kit, Rat Gluc Lung squamous cell carcin Breast invasive ductal ca Cervix squamous cell carc EnzyChrom™ Kinase Assay Esophagus squamous cell c Esophagus squamous cell c
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Role of Liposome Size, Surface Charge, and PEGylation on Rheumatoid Arthritis Targeting Therapy.Rheumatoid arthritis (RA) is a chronic, systemic, progressive autoimmune disease. The vascular permeability of inflamed joints in RA makes it a natural candidate for passive targeting, similar to the enhanced permeability and retention (EPR) effect in solid tumors. Thus, various therapeutic drugs have been encapsulated in nanocarriers to achieve longer in vivo circulation times and improve RA targeting. Although liposomes are the most widely used nanocarriers for RA treatment, the effects of physical and chemical characteristics of liposomes, such as particle sizes, surface charge, polyethylene glycol (PEG) chain length, and PEG concentration, on their passive RA targeting effect have not been fully elucidated. Here, we systematically investigated the effects of physical and chemical properties of liposomes on circulation time and conducted preliminary studies on their passive targeting mechanisms. A series of liposomes with different particle sizes (70, 100, 200, and 350 nm), surface charges (positive, negative, slight positive, and slight negative), PEG chain lengths (1, 2, and 5 kDa), and concentrations (5, 10, and 20% w/w of total lipid) were prepared by lipid film dispersion and extrusion. The pharmacokinetics of liposomes with different formulas were evaluated with a fluorescence microplate reader. A collagen-induced arthritis (CIA) mouse model was utilized to mimic RA pathological conditions and to evaluate the targeting and efficacy of liposomes with different properties using a near-infrared fluorescence imaging system. Uptake of fluorescent liposomes by various synovial cells was measured by flow cytometry. The results indicated that liposomes with 100 nm diameter, a slight negative charge, and 10% incorporation of 5 kDa PEG had better in vivo circulation time and inflamed joint targeting than did other liposomes. Dexamethasone (Dex) was encapsulated into optimized liposomes as an active ingredient for RA treatment. Pharmacodynamic studies demonstrated that Dex liposomes could significantly improve the antiarthritic efficacy of Dex in a CIA mouse model of RA. This study also found that the retention mechanism of RA was mainly increased because of the uptake of liposomes by fibroblasts and macrophages in inflamed joints. This study provides a persuasive explanation for passive RA targeting by liposomes and advances our ability to treat RA with nanomedicine.
1158 related Products with: Role of Liposome Size, Surface Charge, and PEGylation on Rheumatoid Arthritis Targeting Therapy.Bcl-2 Oncoprotein; Clone Bcl-2 Oncoprotein; Clone c-erbB-2 Oncoprotein c-erbB-2 Oncoprotein c-erbB-3 Oncoprotein; Cl c-erbB-3 Oncoprotein; Cl c-erbB-2 Oncoprotein; Cl c-erbB-2 Oncoprotein; Cl c-erbB-2 Oncoprotein; Cl c-erbB-2 Oncoprotein; Cl Bcl-2 Oncoprotein; Clone c-erbB-2 Oncoprotein
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Accessible Telemedicine Diagnostics with ELISA in a 3D Printed Pipette Tip.We report herein a novel pipet-based "ELISA in a tip" as a new versatile diagnostic tool featuring better sensitivity, shorter incubation time, accessibility, and low sample and reagent volumes compared to traditional ELISA. Capture and analysis of data by a cell phone facilitates electronic delivery of results to health care providers. Pipette tips were designed and 3D printed as adapters to fit most commercial 50-200 μL pipettes. Capture antibodies (Ab) are immobilized on the inner walls of the pipet tip, which serves as the assay compartment where samples and reagents are moved in and out by pipetting. Signals are generated using colorimetric or chemiluminescent (CL) reagents and can be quantified using a cell phone, CCD camera, or plate reader. We utilized pipet-tip ELISA to detect four cancer biomarker proteins with detection limits similar to or lower than microplate ELISAs at 25% assay cost and time. Recoveries of these proteins from spiked human serum were 85-115% or better, depending slightly on detection mode. Using CCD camera quantification of CL with femto-luminol reagent gave limits of detection (LOD) as low as 0.5 pg/mL. Patient samples (13) were assayed for 3 biomarker proteins with results well correlated to conventional ELISA and an established microfluidic electrochemical immunoassay.
2181 related Products with: Accessible Telemedicine Diagnostics with ELISA in a 3D Printed Pipette Tip.Beta Amyloid (42) ELISA K GLP 1 ELISA Kit, Rat Gluc Beta Amyloid (1 40) ELISA Beta Amyloid (40) ELISA K Glucagon ELISA KIT, Rat G Beta Amyloid (1 40) ELISA Rabbit Anti-IAA (Indole-3 Rabbit Anti-Integrin alph Rabbit Anti-TNIP2 ABIN2 T Human intercellular adhes Rat intestinal fatty acid Rat Visceral adipose spec
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Optimisation of the Protocol for the LIVE/DEAD BacLight Bacterial Viability Kit for Rapid Determination of Bacterial Load.Rapid antimicrobial susceptibility testing is needed to reduce prescription of inappropriate antibiotics. A rapid alternative to standard culture-based testing is to determine reductions in cell viability using the LIVE/DEAD BacLight Bacterial Viability Kit. We optimised the kit protocol for this application, focusing on simplifying the process by minimising the steps involved and on determining the optimal analytical parameters for fluorescence measurements from the dyes SYTO 9 and propidium iodide (PI). We demonstrate that for our experimental system, the intensity of emissions should be integrated from 505-515 nm for SYTO 9 and 600-610 nm for PI, and the proportion of live cells calculated from a new dye ratio formula, termed the adjusted dye ratio. We show that the pre-staining washing step is not necessary if a non-fluorescent growth media is used; however, staining must be done for each sampling as prolonged exposure to the dyes negatively impacts cell viability. The optimised methodology was able to reproducibly detect reductions in culture viability when the proportion of live cells in a sample of 1 × 10 cells/ml fell below ∼50% live in a media that supports the growth required for detecting antibiotic killing. Finally, we show that the interaction of fluorescence emission spectra from SYTO 9 and PI stained cells is influenced by the proportion of dead cells in a sample. The excitation of PI by SYTO 9 was found to occur in populations containing sufficient numbers of dead cells (>25%), whereas in populations with low numbers of dead cells the dye interaction was additive in regard to red emissions, indicating that these dye interactions may offer another dimension to live/dead analysis. Fluorescence measurements from samples established according to the optimised protocol can be taken using a flow cytometer, spectrofluorometer, microplate reader, and the Optrode, a fibre-based spectroscopic system developed at the University of Auckland.
2122 related Products with: Optimisation of the Protocol for the LIVE/DEAD BacLight Bacterial Viability Kit for Rapid Determination of Bacterial Load.MOUSE ANTI BORRELIA BURGD Live Bacterial Gram Stain Live Bacterial Gram Stain MyGenie 96 Gradient Therm QuantiChrom™ Formaldehy MarkerGeneTM in vivo lacZ MarkerGene™ LysoLive™ Cellufine Formyl , 50 ml Cellufine Formyl Media Cellufine Formyl , 500 ml Cellufine Formyl Media Cellufine Formyl Media
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U13 → C13 mutation in the variable region of the NA gene 3'UTR of H9N2 influenza virus influences the replication and transcription of NA and enhances virus infectivity.The untranslated regions within viral segments are the essential promoter elements required for the initiation of viral replication and transcription. The end of the UTR sequence and part of the ORF sequence constitute the packaging signal for progeny viruses. To explore the influence of single-point and multi-site joint mutations in the UTR of the NA gene on the viral expression, we select clones with upregulated expression of the reporter gene and analyze their sequence characteristics. Bioinformatics methods were used to analyze polymorphisms in the untranslated region (UTR) of the neuraminidase gene of the H9N2 influenza A virus. Using the RNA polymerase I reporting system with enhanced green fluorescence protein (EGFP) gene as the reporter gene, libraries containing random mutations at sites within the N2 UTR were constructed using random mutagenesis. The mutants were selected from the randomized mutagenesis libraries for the N2-UTR. The N2-UTR-RNA polymerase I fluorescence reporter system was identified by sequencing and transfected into infected MDCK cells. The expression of the reporter EGFP was observed using fluorescence microscopy, and the relative fluorescence intensity was measured using a multifunctional microplate reader to analyze the expression of the reporter gene (EGFP) qualitatively and quantitatively. Herein, an RNA polymerase reporter system was constructed to rescue the mutated viruses and measure their tissue culture infective dose (TCID50). The results showed that the U13 → C13 mutation in the 3'end of the NA gene promoted the expression of viral RNA and protein, and mutation of other sites within the UTR could differentially regulate viral genomic transcription and translation. These data showed that the U13 → C13 mutation within the variable region of the 3'UTR of the NA gene in the H9N2 influenza virus promotes viral genomic expression and infection.
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A high-throughput screen for the identification of compounds that inhibit nematode gene expression by targeting spliced leader trans-splicing.Infections with parasitic nematodes are among the most significant of the neglected tropical diseases affecting about a billion people living mainly in tropical regions with low economic activity. The most effective current strategy to control nematode infections involves large scale treatment programs with anthelmintic drugs. This strategy is at risk from the emergence of drug resistant parasites. Parasitic nematodes also affect livestock, which are treated with the same limited group of anthelmintic drugs. Livestock parasites resistant to single drugs, and even multi-drug resistant parasites, are appearing in many areas. There is therefore a pressing need for new anthelmintic drugs. Here we use the nematode Caenorhabditis elegans as a model for parasitic nematodes and demonstrate that sinefungin, a competitive inhibitor of methyltransferases, causes a delay in development and reduced fecundity, and inhibits spliced leader trans-splicing. Spliced leader trans-splicing is an essential step in gene expression that does not occur in the hosts of parasitic nematodes, and is therefore a potential target for new anthelmintic drugs. We have exploited the ability of sinefungin to inhibit spliced leader trans-splicing to adapt a green fluorescent protein based reporter gene assay that monitors spliced leader trans-splicing for high-throughput screening for new anthelmintic compounds. We have established a protocol for robust high-throughput screening, combining mechanical dispensing of living C. elegans into 384- or 1536- well plates with addition of compounds using an acoustic liquid dispenser, and the detection of the inhibition of SL trans-splicing using a microplate reader. We have tested this protocol in a first pilot screen and envisage that this assay will be a valuable tool in the search for new anthelmintic drugs.
1043 related Products with: A high-throughput screen for the identification of compounds that inhibit nematode gene expression by targeting spliced leader trans-splicing.EnzyChrom™ Kinase Assay DNA (cytosine 5) methyltr Mouse L-308 Array, Glass Mouse L-308 Array, Glass Mouse L-308 Array, Membra Mouse L-308 Array, Membra Rat L-90 Array, Glass Sli Rat L-90 Array, Glass Sli Rat L-90 Array, Membrane Rat L-90 Array, Membrane Gene Expression: Mouse N Gene Expression: Rat P45
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A reliable fluorimetric method to screen the nitric oxide synthase inhibitors in 96 well plate.In general, 4 amino-5-methylamino-2',7'-difluorescein diacetate (DAF-FM-DA) dye is used to detect nitric oxide in biological systems through cell imaging. In this study, we have used 96 well plate format to quantify nitric oxide using DAF-FM-DA through a multimode reader (or independently using fluorospectrometer) and could be visualized in a fluorescence microscope. Similar study otherwise will require a high-end instrument. The method has been validated to screen NOS inhibitors in the HEK 293T cell lines over-expressing the NOS isoforms. We observed that the method is very simple to use, adaptive, sensitive and most importantly it saves time. REAGENTS/TOOLS: Ethanol (70% [v/v] in distilled water), Nω-Nitro-l-arginine (l-NAME), 7-Nitro-Indazole (7-NI) (Sigma, St. Louis, MO), HEK 293T cell lines (National Centre for Cell Science (NCCS), Pune, India), DMEM (Himedia laboratories Pvt), Fetal Bovine Serum (FBS) (Invitrogen, Carlsbad, CA), 100 U/mL penicillin, and 0.1 mg/mL streptomycin in a 5% CO2 atmosphere. Hank's Balanced Salt Solution (HBSS) without Phenol Red of pH 7.4 was prepared with the following composition: NaCl, 8.0g, KCl, 0.4g, CaCl, 0.14g, MgSO⋅7HO, 0.1g, MgCl·6HO, 0.1g, NaHPO·2HO, 0.06g, KHPO, 0.06g, glucose, 1.0g, NaHCO, 0.35g, HO, to 1000 ml, Sterilized and refrigerated, Calcium Ionophore A23187 (Sigma Aldrich 52665-69-7) DAF-FM Di Acetate (Molecular Probes Life Technologies), and DAF-FM Di Aceatate was prepared as a stock solution (5 mM) in DMSO, divided into aliquots and stored at -20 °C, followed by dilution to the required concentration in HBSS buffer before use. EQUIPMENT: Neubauer chamber, Microtube centrifuges (1.5 mL), Micropipettors,10,100, and 1000 mL with corresponding tips, multimode reader (Tecan, Synergy-HT), inverted fluorescence microscope (Nikon, eclipse Ti-S), black flat bottom Microplates (96-well) (Corning 3603).
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[Effects of sulfur dioxide on alveolar macrophage apoptosis in acute lung injury induced by limb ischemia/reperfusion in rats].To investigate the effect of sulfur dioxide (SO) on the apoptosis of alveolar macrophage (AM) in lung protection of limb ischemia/reperfusion (I/R) induced acute lung injury (ALI), and to find a new target for the control of inflammatory response.
2917 related Products with: [Effects of sulfur dioxide on alveolar macrophage apoptosis in acute lung injury induced by limb ischemia/reperfusion in rats].Rabbit Anti-Human Apoptos Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Anti AGO2 Human, Monoclon Anti AGO2 Mouse, Monoclon Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Macrophage Inflamma Human Epstein-Barr Virus Human Gro g Macrophage In
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