Three tetrazolium salts are used as histochemical indicators of dehydrogenase activity in the skin of sheep and other mammals. Of these compounds 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyl-tetrazolium chloride (INT) is shown to be the most satisfactory. It enables various histological structures of the wool follicle to be conveniently demonstrated.. In the presence of appropriate substrates and DPN and ATP, the intracellular deposition of the granular formazan in frozen sections has revealed the distribution of succinic, α-glycerophosphoric, lactic, and malic dehydrogenases, and regions where some phosphorylated carbohydrate intermediates promote reduction. Metabolic activity in the Malpighian layers and in the undifferentiated epidermal cells of the hair matrix reduces INT enzymatically up to the pre-keratinous zone. Here the protein-bound sulphydryl groups reduce INT and produce intense red staining which is easily distinguished from the granular enzymic staining.. The distribution of ...
I know Dr Mike Wilson at the Insitute for Dental Research, 256 Grays Inn Road, London knows about this. It might be worth contacting him. Julian Wimpenny, sabjw at Cardiff.ac.uk ...
Since heterotrophic prokaryotes play an important biogeochemical role in aquatic ecosystems and have a high capacity to survive in extreme environments, easy-to-perform protocols that probe their physiological states and the effects of environmental variables on those states are highly desired. Some methodologies combine a general nucleic acid stain with a membrane integrity probe. We calibrated one of these, the nucleic acid double-staining (NADS) protocol (G. Gre ́gori, S. Citterio, A. Ghiani, M. Labra, S. Sgorbati, S. Brown, and M. Denis, Appl. Environ. Microbiol. 67:4662-4670, 2001), determining the optimal stain concentrations in seawater and the response to conditions that generate prokaryote death (such as heat) and to conditions that are known to produce death in plankton, such as nutrient limitation or flagellate grazing. The protocol was validated by comparison to two methods used to detect viability: active respiration by 5-cyano 2,3-ditolyl tetrazolium chloride (CTC) and ...
For studying proliferation and determination of survival of cancer cells after irradiation, the multiple MTT assay, based on the reduction of a yellow water soluble tetrazolium salt to a purple water...
Measurement of cell proliferation is necessary for testing the effects of pharmacological agents or growth factors, assessing cytotoxicity or investigating circumstances of cell activation. In a cell proliferation assay, the increase in number of cells or change in the proportion of cells that is dividing is assessed. Trevigen® offers a Calcein-AM based kit to determine cell viability as well as the tetrazolium salts MTT and XTT metabolic cell proliferation assay kits.. ...
Various assays, using different strategies, are available for assessing cultured cell proliferation. These include measurement of metabolic activity (tetrazolium salts and alamarBlue), DNA quantification using fluorophores (Hoechst 33258 and PicoGreen), uptake of radioactively-labeled DNA precursors such as [3H]thymidine, and physical counting (hemocytometer). These assays are well established in characterizing cell proliferation in two-dimensional (2D), monolayer cultures of low cell densities. However, increasing interest in 3D cultures has prompted the need to evaluate the effectiveness of using these assays in high cell density or 3D cultures. We show here that typical cell proliferation assays do not necessarily correlate linearly with increasing cell densities or between 2D and 3D cultures, and are either not suitable or only rough approximations in quantifying actual cell numbers in a culture. Prudent choice of techniques and careful interpretation of data are therefore recommended when ...
Massoumi, S. "PMN inactivation of Candida albicans-MTT-reduction : a three hour model system suggesting that MTT reduction detects dead cells" (1992). OHSU Digital Collections. https://doi.org/10.6083/M4PN93QN ...
Utilizes the second generation tetrazolium dye, XTT (sodium 2,3,-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)-carbonyl]-2H-tetrazolium inner salt), to measure cell growth, cytotoxicity, and apoptosis. It is a homogenous, highly sensitive, reproducible assay, and adaptable to high-throughput screening.|br />
A. W. Segal, A. J. Levi; The Mechanism of the NBT (Nitroblue Tetrazolium) Test. Clin Sci Mol Med 1 June 1973; 44 (6): 26P. doi: https://doi.org/10.1042/cs044026Pa. Download citation file:. ...
A stable reagent for assaying an analyte from a fluid sample which includes an enzyme, a mediator, a tetrazolium salt and an oxidizing agent selected from the group consisting of sodium chlorate, 2,5-dimethylhexane-2,5-dihydroperoxide, benzoyl peroxide, t-butyl peroxide, sodium iodate, N-ethylmaleimide, t-butylperoxyacetate, nickel acetylacetonate, stannic chloride, rhodium (III) trichloride hydrate, and t-butylperbenzoate is described. Inclusion of an oxidizing agent from the group specified above stabilizes the reagent, thereby preventing a high blank reaction because a high blank introduces error into the assay of an analyte. The reagent may be provided in solution form by adding water, or may be included in a film by adding a film-forming agent. The reagent may be utilized to perform the assay of an analyte from a fluid sample. The assay may be conducted in a solution medium by adding the fluid sample to the liquid reagent, and spectrophotometrically measuring absorbance of the color indicating the
Rapid viability tests of the Category B agent Escherichia coli O157:H7 were evaluated after disinfection with chlorine. The metabolic activity dyes ChemChrome V6, a modified fluorescein diacetate (FDA) and 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) were compared to standard plate counts. ChemChrome V6 results were obtained using a solid phase cytometer and CTC results by microscopic analysis. The water-borne bacteria Legionella pneumophila and Mycobacterium avium were also tested as positive and negative controls. Under the conditions tested, CTC provided more consistency with plate count estimates of viability than did ChemChrome V6. ...
Peer reviewed online journal covering broad area in biomedical engineering All free for submission, publication, and download The official journal of Japanese Society for Medical and Biological Engineering (JSMBE)
The glucose content of blood can be determined by contacting a blood sample with a test strip containing a glucose dehydrogenase dependent on PQQ (or derivatives or isomers) thereof as a co-factor, a tetrazolium salt indicator, but in the absence of a mediator.
3, 3-(3, 3-Dimethoxy[1, 1-biphenyl]-4, 4-diyl)-bis(2, 5-diphenyl-2H-tetrazolium) dichloride; 3, 3-Dianisole-4, 4-bis(3, 5-diphenyltetrazolium chloride); BTC; Blue Tetrazolium chloride; ...
A colorimetric cell proliferation assay using soluble tetrazolium sodium [(CellTiter 96? Aqueous One Remedy) cell proliferation reagent, comprising the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) and an electron coupling reagent phenazine ethosulfate], was optimized and certified for quantitative dedication of IL-15 dependent CTLL-2 cell proliferation activity. of scripts written in the R Statistical Language and Environment utilizing a […]. ...
* found in: Thiazoyl Blue Tetrazolium Bromide (MTT), 4-MUG (4-Methylumbelliferyl-beta-D, Quasar DX™ Soluble/ELISA and Membrane Blotting Chemiluminescent..
Doctors have a new ally to deal with cases of cancer patients who have two or more possible treatments: the chemosensitivity test.
WST-1 premixed Cell Proliferation Assay Kit- An effortless protocol, start to finish under 4 hours. Get 2,500 Rxns for $316.00. Skip washing steps and additional reagents.
WST-1 premixed Cell Proliferation Assay Kit- An effortless protocol, start to finish under 4 hours. Get 2,500 Rxns for $316.00. Skip washing steps and additional reagents.
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Introduction In the present study, confocal microscopy, a miniflow cell system, and image analysis were combined to test in situ the effect of antimicrobials and alkali on biofilms of Enterococcus faecalis, Lactobacillus paracasei, Streptococcus anginosus, and Streptococcus gordonii isolated from root canals with persistent infections. Methods Biofilms formed for 24 hours were exposed for 5 minutes to alkali (pH = 12), chlorhexidine digluconate (2.5%), EDTA (50 mmol/L), and sodium hypochlorite (1%). The biofilms were then characterized by using fluorescent markers targeting cell membrane integrity (LIVE/DEAD) and metabolic activity (5-cyano-2,3-ditolyl tetrazolium chloride and fluorescein diacetate). In addition, the biofilm architecture and the extent to which coating of the substrate surface with collagen influenced the resistance pattern to the chemicals were also analyzed. Results NaOCl (1%) affected the membrane integrity of all organisms and removed most biofilm cells. Exposure to EDTA (50 ...
IQBAL, M. et al. Assessment of Nili-Ravi buffalo (Bubalus bubalis) semen by MTT reduction assay. S. Afr. j. anim. sci. [online]. 2009, vol.39, n.5, pp.294-300. ISSN 2221-4062.. MTT (3-(4,5-dimethyl thiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay is commonly used to validate the viability of metabolically active cells. The study was conducted to examine and validate the MTT test to assess the sperm viability of Nili-Ravi buffalo bulls and compare the efficiency of the test with the supra-vital staining technique (eosin-nigrosine) and hypo-osmotic swelling test. Fresh semen samples from breeding Nili-Ravi buffalo bulls (n = 20) were collected using an artificial vagina. After assessing the quality of semen for normal parameters, the MTT assay was carried out in phosphate buffer saline. Results revealed a high significant correlation (r = 0.995) between the viability of sperm and the rate of reduction of MTT. The other proportions of some semen samples showed a weak relationship between the ...
Looking for online definition of NBT test in the Medical Dictionary? NBT test explanation free. What is NBT test? Meaning of NBT test medical term. What does NBT test mean?
Find quality suppliers and manufacturers of 4727-29-1(Benzoic acid,2-[(phenylamino)carbonyl]-) for price inquiry. where to buy 4727-29-1(Benzoic acid,2-[(phenylamino)carbonyl]-).Also offer free database of 4727-29-1(Benzoic acid,2-[(phenylamino)carbonyl]-) including MSDS sheet(poisoning, toxicity, hazards and safety),chemical properties,Formula, density and structure, solution etc.
Purpose: : Oxidative damage to the Retinal Pigment Epithelium (RPE) is thought to play a major role in the development and progression of Age-related Macular Degeneration (AMD). The purpose of this in vitro study was to evaluate the ability of the selective 5-HT1A agonist, AL-8309A, to protect RPE cells against oxidative damage and to define the mechanism of the cytoprotective action. Methods: : Using the human RPE cell line ARPE-19, we have developed an in vitro survival assay using the cytotoxin 7-ketocholesterol (7kCh). Oxidative damage and apoptosis is induced by the addition of 7kCh, and the tetrazolium salt, WST-1, is used to assess cell viability following treatment. AL-8309A was evaluated in this model at concentrations from 0.01 nM to 100 µM. Western analysis was performed to assess changes in protein concentration and activation state in response to treatment with AL-8309A. The selective 5-HT1A antagonists, NAN-190 and WAY 100635, and specific Mek inhibitor, U0126, were used to ...
This study investigated the interaction between eupatorin and bovine serum albumin (BSA) using ultraviolet-visible (UV-vis) absorption, fluorescence, synchronous fluorescence, circular dichroism (CD) spectroscopies, and molecular modeling at pH 7.4. Results of UV-vis and fluorescence spectroscopies illustrated that BSA fluorescence was quenched by eupatorin via a static quenching mechanism. Thermodynamic parameters revealed that hydrophobic and electrostatic interactions played major roles in the interaction. Moreover, the efficiency of energy transfer, and the distance between BSA and acceptor eupatorin, were calculated. The effects of eupatorin on the BSA conformation were analyzed using UV-vis, CD, and synchronous fluorescence. Finally, the binding of eupatorin to BSA was modeled using the molecular docking method.
Particularly preferably, Z21 is 2,4-dichloro-1,3,5-triazin-6-ylf 2-chloro-4-(3-(2-suifatoethylsulfonyl)-phenylamino)-1,3,5-triazin-6-yl, 2-chloro-4-(4-(2-su!fatoethy!sulfonyl)-phenylamino)-1,3,5-triazin-6-yl, 2-chloro-4-(3-(vinylsulfonyl)-phenylarnino)-1 t3,5-triazin-6-yl, 2-chloro4-(4-(vinylsulfonyl)-phenylamino)-1,3,5-triazin-6-yl, 2-ch!oro-4-(N-methy!-N-(2-(2-sulfatoethylsulfonyl)-ethy!)-amino)-1,3,5-triazin-6-yl, 2-ch!oro-4-(N-phenyl-N-(2-(2-sulfatoethylsulfonyl)-ethyl)-amino)-1,3,5-tria2in-6-yl, 2-fluoro-4-morpholino-1,3,5-triazin-6-yI, 2-fluoro-4-(2-sulfophenyl-amino)-1,3,5-triazin-6-yl, 2-fluoro-4-(3-sulfophenylamino)-1,3,5-triazin-6-yl, 2-fluoro-4-(4-sulfophenylamino)-1,3,5-triazin-6-yl, 2-fiuoro-4-(3-trimethylammonio-phenylamino)-1,3,5-triazin-6-ylf 2-fIuoro-4-(4-trirnethylammoniophenylamino)-1,3,5-triazin-6-yl, 2-fluoro-4-(3-(2-sulfatoethylsulfonyl)-phenylamino)-1,3,5-triazin-6-yl( 2-fluoro-4-(4-(2-sulfatoethylsulfonyl,-phenylamino)-1 f3,5-triazin-6-yl, ...
With the aim of improved prediction of human skin irritation, cytotoxicity tests were conducted using a reconstituted human dermal model (Skin2TM cultured 3-Dimensional Skin Model, ZK1300: 3-D model). A range of water-insoluble substances, namely fatty acids, fatty alcohols and hydrocarbons, were tested with MTT(3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyl tetrazolium bromide) reduction, LDH (lactate dehydrogenase) release and prostaglandin E2(PGE2) release using as cytotoxicity markers. Results for these parameters were compared with those obtained from a cytotoxicity test (MTT reduction assay) using cultured monolayers of human dermal fibroblasts (NB1RGB), from a rabbit primary skin irritation test and from human patch testing ...
METHODS AND RESULTS Twenty-one dogs with 3 hours of left anterior descending coronary artery occlusion and 2 to 3 hours of reflow were studied. MCE was performed at 15 and 45 minutes and 2 and 3 hours after reflow. It was also performed at either 2 or 3 hours after reflow in the presence of 0.56 mg/kg of dipyridamole. Radiolabeled microsphere-derived blood flow was measured at 15 minutes and 2 and 3 hours after reflow and during dipyridamole effect. Infarct size was measured at the end of the experiment by use of triphenyl tetrazolium chloride. MCE data were processed with color-coding schemes that highlighted differences in myocardial videointensities in proportion to the concentration of microbubbles within the microvasculature. There was significant variability in MCE-defined perfusion patterns after reflow, with contrast defects noted mainly within the endocardium. There were fair and significant (P , .05) correlation (r = -.73 to r = -.55) between MCE defect size and normalized endocardial ...
2-{[5-Fluoro-2-(phenylamino)pyrimidin-4-yl]amino}benzoic acid | C17H13FN4O2 | CID 23647269 - structure, chemical names, physical and chemical properties, classification, patents, literature, biological activities, safety/hazards/toxicity information, supplier lists, and more.
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
Cell culture and toxicity studies. The HT-22 hippocampal nerve cell line is a subclone of HT4 (Morimoto and Koshland, 1990). The HT-22 clone was selected for its sensitivity to glutamate toxicity. The cells do not possess active ionotropic glutamate receptors and are not subject to excitotoxicity (Maher and Davis, 1996). HT-22 cells are propagated in DMEM (Vogt and Dulbecco, 1963) supplemented with 10% fetal bovine serum. Cell survival was determined by the MTT [3-(4,5-dimethyldiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assay as described (Schubert et al., 1992), which in this cell system, correlates with cell death as determined by trypan blue exclusion and a colony-forming assay (Davis and Maher, 1994). Briefly, HT-22 cells are dissociated with pancreatin (Life Technologies, Gaithersburg, MD) and seeded onto 96-well microtiter plates in 5% dialyzed fetal bovine serum at a density of 2.5 × 103 cells per well in 100 μl of medium. The next day cells are treated with various reagents ...
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4BCK: Comparative Structural and Functional Studies of 4-(Thiazol- 5-Yl)-2-(Phenylamino)Pyrimidine-5-Carbonitrile Cdk9 Inhibitors Suggest the Basis for Isotype Selectivity.
4BCF: Comparative Structural and Functional Studies of 4-(Thiazol- 5-Yl)-2-(Phenylamino)Pyrimidine-5-Carbonitrile Cdk9 Inhibitors Suggest the Basis for Isotype Selectivity.
The MTT assay is based on the conversion of MTT into formazan crystals by living cells, that determines number of viable cells. The treatment of K562 cells with different concentrations of metylthiosemicarbazon complex with Zn2+ (30, 50, 80, 100, 130, 150 and 200µM) at 24, 48 and 72 h. As showen Fig, metylthiosemicarbazon complex with Zn2+ has cytotoxic activity on the K562 cell line and was able to inhibit the proliferation of the K562 cell line. The IC50 is a measure of the effectiveness of metylthiosemicarbazon complex with Zn2+ in inhibiting of proliferation of K562 cells. The IC50 value of this compound is 100µM in 72 h.. Figure 1: Effect of metylthiosemicarbazon complex with Zn2+ on K562 cells. The K562 cells were treated with various concentrations (30-150µM) of metylthiosemicarbazon complex with Zn2+ for 24, 48, 72 h and then were investigated by MTT assay. Data are shown as mean ± SD.. 3.2 Morphological study of K562 ...
The previously described 2-aryl-4-chloro-3-iodoquinolines were prepared following literature procedure and in turn converted to the corresponding hitherto unknown 2-aryl-3-iodo-4-(phenylamino)quinoline derivatives using aniline in refluxing ethanol. These 2-aryl-3-iodo-4-(phenylamino)quinolines were reacted with allybromide in ethanol at room temperature to afford 4-(N,N-allylphenylamino)-2-aryl-3-iodoquinoline derivatives. The 2-aryl-3-iodo-4-(phenylamino)quinoline and 4-(N,N-allylphenylamino)-2-aryl-3-iodoquinoline derivatives were subjected to metal-catalysed carbon-carbon bond formations. Palladium(0)-copper iodide catalysed Sonogashira cross-coupling of 2-aryl-3-iodo-4-(phenylamino)quinoline with terminal alkynes afforded series of 1,2,4-trisubstituted 1H-pyrrolo[3,2-c]quinolines in a single step operation. On the other hand, the 4-(N,N-allylphenylamino)-2-aryl-3-iodoquinoline derivatives were found to undergo palladium-catalysed intramolecular Heck reaction to yield the corresponding ...
This in vitro study evaluated the influence of chlorhexidine diacetate (CDA) when blended within dentin bonding systems (DBSs) on Streptococcus mutans (S. mutans) biofilm formation. One commercially available 0.2% wt CDA-containing DBS (Peak Universal Bond) and five experimental 0.2% wt CDA-containing DBS formulations (experimental Adper Scotchbond 1XT plus experimental resins, R2, R3, R4, R5) were assessed vs their no-CDA containing counterparts. Twenty-eight DBSs disks were prepared for each group (6.4 mm×1.0 mm) and cured for 80 s at 800 mW/cm2 in a nitrogen atmosphere. A modified Drip-Flow Reactor was used to grow S. mutans biofilms on specimen surfaces for 24 h and adherent, viable biomass was evaluated using a tetrazolium salt assay (MTT). Two specimens from each of the tested materials were processed with LIVE/DEAD stain and observed using laser confocal microscopy (CLSM) while two disks from each group were examined by using scanning electron microscopy (SEM). MTT assay, CLSM and SEM ...
RECENT FINDINGS: R. seeberi, the pathogen that causes rhinosporidiosis, has been definitively classified using molecular biological tools in a new clade - the Mesomycetozoea, along with 10 parasitic and saprobic microbes. The controversial spherical bodies of the endospores have been shown to comprise both lipid/protein nutritive bodies and other spherical bodies that are metabolizing units that reduce MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide). This indicates the viability of these spherical bodies, provisionally identified as the electron dense bodies that have also been shown to contain nucleic acids. MTT reduction as an indicator of viability has been used to determine the sensitivity of rhinosporidial endospores to biocides, antimicrobial drugs, and to specific antibodies. Genetic heterogeneity has been identified in strains from humans and animals. Cell-mediated and humoral immune responses have been demonstrated in human patients and in mice. Several mechanisms ...
RECENT FINDINGS: R. seeberi, the pathogen that causes rhinosporidiosis, has been definitively classified using molecular biological tools in a new clade - the Mesomycetozoea, along with 10 parasitic and saprobic microbes. The controversial spherical bodies of the endospores have been shown to comprise both lipid/protein nutritive bodies and other spherical bodies that are metabolizing units that reduce MTT (3-[4,5-dimethyl-2-thiazolyl]-2,5-diphenyl-2H-tetrazolium bromide). This indicates the viability of these spherical bodies, provisionally identified as the electron dense bodies that have also been shown to contain nucleic acids. MTT reduction as an indicator of viability has been used to determine the sensitivity of rhinosporidial endospores to biocides, antimicrobial drugs, and to specific antibodies. Genetic heterogeneity has been identified in strains from humans and animals. Cell-mediated and humoral immune responses have been demonstrated in human patients and in mice. Several mechanisms ...
African Traditional Medicines (ATMs) serve as a major source of primary healthcare for African people. The reasons for their use range from easy access, affordability, beliefs in traditional systems and long term safety. ATMs have been used to treat individuals infected with HIV and therefore need scientific validation; a view supported by Traditional Health Practitioners (THPs). This study aimed to evaluate the in vitro cytotoxicity, immune modulatory and anti-HIV activities of traditional multiple herbal preparations from local THPs. Ugambu, Ihashi, Product Nene, Product Blue, SPNa and SDKc ATM were supplied by local THPs. Changes in adenosine triphosphate (ATP) & glutathione (GSH) over 24 hours were measured using luminometry. Changes in 12 cytokines were assayed using an ELISA-based absorbance assay. Protective effects against HIV killing of MT-4 cells were tested using the XTT assay and antiviral activity was measured using an HIV-1 viral load assay. Cyclosporine and AZT were used as ...
SIM deeps are a multi-test medium comprising 3 tests: sulfide (H2S gas), indole production, and motility. You have already tested for motility via the hanging drop slide, and here is an additional way to determine it, with less muss and fuss. In fact, there is an even better way to determine motility, motility agar with tetrazolium dye, that you will also be using in lab. Both of these agars have advantages. SIM is actually 3 tests in one, but, on the other hand, the motility with tetrazolium is much easier to read for motility. The tetrazolium is a colorless salt which becomes red when reduced, occurring as a result of bacterial metabolism.. ...
Aim Rosa damascene Mill. belongs to the family of Roseaceae and its essential oil is produced in large amounts in Iran. The wide application of rose oil has raised questions about potential adverse health effects. We have investigated cytotoxic activity and genotoxic effects of Rosa oil from Kashan, Iran. Methods The cytotoxic effect and IC50 of the essential oil on the cell lines was studied followed by MTT assay. In this assay mitochondrial oxidoreductase enzymes with reducing the tetrazolium dye MTT (3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide) reflect the number of viable cells ...
The aim of this study was to use an in vitro (3-(4,5-dimethylthiazol-2-yl)-5-(3- carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) (MTS) assay in combination with an explant culture technique to determine ...
Cell lines. The HNSCC cell lines CAL27 and UM-SCC1, representing oral cavity carcinomas, were used. CAL27 was obtained from the American Type Culture Collection, and the UM-SCC1 cell line was obtained from Dr. Thomas E. Carey (University of Michigan, Ann Arbor, MI). Cell lines were grown in DMEM containing high glucose (4,500 μg/mL) and 1 mmol/L of glutamine, 100 IU/mL penicillin, 100 IU/mL streptomycin, 0.5 μg/mL fungizone, and 10% fetal bovine serum (Sigma).. Cell viability. Growth medium was aspirated out of wells, taking care not to disturb the residual cell mass at the bottom of the plate. Following this, 0.5 mL of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution (1 mg/mL in complete medium; Sigma) was added to each well, followed by incubation at 37°C for 1 h until the solution turned purple. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide solution was aspirated out of the wells and air-dried for 5 min. Serial dilution of the cells was done in ...
Cytotoxicity assay. Cell viability was assessed with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (21). Cells were seeded into the 96-well microplates at 1 × 104 per well followed by IG-105 treatment at concentrations between 0 and 1 μmol/L for 72 h at 37°C. The supernatants were removed gently and cell viability was assessed. The plate was read in a microplate reader. The IC50 was determined in duplicates and each experiment was repeated at least three times under identical conditions. IC50 value was defined as the drug concentration that inhibits 50% cell growth compared with the untreated controls and calculated by regression analysis.. Pgp ATPase assay. Drug-stimulated activity of Pgp ATPase was detected by Pgp-Glo assay system (Promega). By following the user protocol provided by the vender, the activity of Pgp ATPase was measured in the presence or absence of 100 μmol/L Na3VO4, 200 μmol/L Verapamil (as a positive reference), 2 μmol/L vincristine, or 2 ...
... , B. Kaskatepe, M. E. Kiymaci, D. Simsek, H. B. Erol and S. A. Erdem1
Peptides , Amyloid Peptides , Beta-Amyloid (1-40) and Related Peptides , Beta-Amyloid (1-40) HCl, Human; All the HCl salt forms of A (1-40), A (25-35) and D-Ser26 A (25-35), take the -structure within a few hours in PBS. They form fibrils, exert toxic effects on hippocampal cultured neurons and suppress MTT reduction activity of non-neuronal (HeLa) cells without cytotoxicity.; DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVV HCl; H-Asp-Ala-Glu-Phe-Arg-His-Asp-Ser-Gly-Tyr-Glu-Val-His-His-Gln-Lys-Leu-Val-Phe-Phe-Ala-Glu-Asp-Val-Gly-Ser-Asn-Lys-Gly-Ala-Ile-Ile-Gly-Leu-Met-Val-Gly-Gly-Val-Val-OH HCl