scholarly journals Advantages and Limitations of Salmon-Gal/Tetrazolium Salt Histochemistry for the Detection of LacZ Reporter Gene Activity in Murine Epithelial Tissue

2017 ◽  
Vol 65 (4) ◽  
pp. 197-206 ◽  
Author(s):  
Claudia Merkwitz ◽  
Orest Blaschuk ◽  
Jana Winkler ◽  
Angela Schulz ◽  
Simone Prömel ◽  
...  
Keyword(s):  
Epithelial Tissue ◽  
Tetrazolium Salt ◽  
Potassium Iodide Solution ◽  
Lacz Gene ◽  
Tetrazolium Chloride ◽  
Blue Tetrazolium Chloride ◽  
Iodide Solution ◽  
Filiform Papillae ◽  
Iodonitrotetrazolium Chloride ◽  
Nbt Staining

The Escherichia coli LacZ gene is a widely used reporter for gene regulation studies in transgenic mice. It encodes bacterial β-galactosidase (Bact β-Gal), which causes insoluble precipitates when exposed to chromogenic homologues of galactose. We and others have recently reported that Bact β-Gal detection with Salmon-Gal (S-Gal) in combination with nitro blue tetrazolium chloride (NBT) is very sensitive and not prone to interference by acidic endogenous β-galactosidases. Unfortunately, as we show here, the method appears to be inadequate for evaluation of Bact β-Gal expression in keratinized epithelial appendages but not in other keratinized epithelia. NBT in the reaction mixture, just as other tetrazolium salts, inevitably causes unwanted staining artifacts in lingual filiform papillae, penile spines, and hair fibers by interacting with keratin sulfhydryl-rich regions. The methodological limitation can be overcome in part by pretreating the tissues before the S-Gal/NBT staining with an iodine–potassium iodide solution. Alternatively, the use of iodonitrotetrazolium chloride instead of NBT in the S-Gal reaction mixture provides enough color resolution to distinguish the specific Bact β-Gal staining in orange from the artifact staining in dark red. In summary, we provide evidence that S-Gal/NBT histochemistry has limitations, when staining keratinized epithelial appendages.

scholarly journals Phenylmethoxybis(tetrazolium) ion-association complexes with an anionic indium(III) — 4-(2-pyridylazo)resorcinol chelate

2013 ◽  
Vol 11 (2) ◽  
pp. 280-289 ◽  
Author(s):  
Teodora Stefanova ◽  
Kiril Gavazov
Keyword(s):  
Limit Of Detection ◽  
Ion Association ◽  
Molar Absorptivity ◽  
Tetrazolium Salt ◽  
Limit Of Quantification ◽  
Tetrazolium Chloride ◽  
Liquid Liquid Extraction ◽  
Blue Tetrazolium ◽  
Blue Tetrazolium Chloride ◽  
Molar Absorptivity Coefficient

AbstractComplex formation and liquid-liquid extraction were studied in systems containing indium(III), 4-(2-pyridylazo)resorcinol (PAR), phenylmethoxybis(tetrazolium) salt (MBT), water and chloroform. The following MBTs, which differ only by the number of -NO2 groups in their cationic parts, were used: 3,3′-(3,3′-dimetoxy-4,4′-biphenylene)bis(2,5-diphenyl-2H-tetrazolium chloride) (Blue Tetrazolium chloride, BT), 3,3′-(3,3′-dimetoxy-4,4′-biphenylene)bis[2-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride] (Nitro Blue Tetrazolium chloride, NBT) and 3,3′-(3,3′-dimetoxy-4,4′-biphenylene)bis[2,5-di(4-nitrophenyl)-2H-tetrazolium chloride] (Tetranitro Blue Tetrazolium chloride, TNBT). The composition of the formed ternary complexes was determined, In:PAR:MBT=1:2:2, and the optimum conditions for their extraction found: pH, shaking time, concentration of the reagents and the sequence of their addition. Some key constants were estimated: constants of extraction (Kex), constants of association (β) and constants of distribution (KD). BT appears to be the best MBT for extraction of the In(III)-PAR species, [In3+(OH)3(PAR)2]4−, (Log Kex=10.9, Log β=9.8, Log KD=1.12, R%=92.7%). Several additional characteristics concerning its application as extraction-spectrophotometric reagent were calculated: limit of detection (LOD = 0.12 µg cm−3), limit of quantification (LOD = 0.40 µg cm−3) and Sandell’s sensitivity (SS =1.58 ng cm−2); Beer’s law is obeyed for In(III) concentrations up to 3.2 µg mL−1 with a molar absorptivity coefficient of 7.3×104 L mol−1 cm−1 at λmax=515 nm.

scholarly journals The solvent extraction and spectrophotometric determination of vanadium (V) with 4-(2-thiazolylazo)resorcinol and tetrazolium salts

2007 ◽  
Vol 5 (1) ◽  
pp. 257-270 ◽  
Author(s):  
Kiril Gavazov ◽  
Vanya Lekova ◽  
Atanas Dimitrov ◽  
Georgi Patronov
Keyword(s):  
Association Constants ◽  
Tetrazolium Salt ◽  
Fast Method ◽  
Tetrazolium Chloride ◽  
Blue Tetrazolium Chloride ◽  
Tetrazolium Violet ◽  
Ion Associate ◽  
The Relationship ◽  
Optimum Extraction

AbstractThe formation and extraction of ion-associate complexes between the vanadium(V)-4-(2-thiazolylazo)resorcinol (TAR) anionic chelate and the cations of some mono-and ditetrazolium salts {3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H-tetrazolium bromide (Thiazolyl blue, MTT), 3-(2-naphtyl)-2,5-diphenyl-2H-tetrazolium chloride (Tetrazolium violet), 2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride (Iodonitrotetrazolium chloride), 3,3′-[3,3′-dimetoxy(1,1′-biphenyl)-4,4′-diyl]-bis[2,5-diphenyl-2H-tetrazolium] chloride (Tetrazolium blue chloride) and 3,3′-(3,3′-dimetoxy-4,4′-biphenylene)bis[2-(4-nitrophenyl)-5-phenyl-2H-tetrazolium chloride] (Nitro blue tetrazolium chloride)} have been studied. The optimum extraction conditions have been found. The composition of the V-TAR-monotetrazolium and V-TAR-ditetrazolium complexes extracted into chloroform has been determined to be 1:2:3 and 2:4:3 respectively. The extraction, distribution and association constants, and the recovery factors have been calculated. The relationship between the molecular weight of tetrazolium cations, and the association constants of their complexes has been discussed. The special behavior of the tetrazolium cations, containing-NO2 groups has been noticed. The effects of foreign ions and reagents on the extraction of vanadium with TAR and the best tetrazolium salt-MTT have been studied. A sensitive, selective, simple and fast method for the determination of vanadium has been developed.

Porcine Repeat Element DNA: In Situ Detection of Xenotransplanted Cells

1995 ◽  
Vol 4 (2) ◽  
pp. 253-256 ◽  
Author(s):  
Henry F. Oettinger ◽  
Amelie Rodrigue-Way ◽  
Joyce J. Bousquet ◽  
Albert S.B. Edge
Keyword(s):  
Southern Blotting ◽  
Host Tissue ◽  
Repeat Element ◽  
Tissue Analysis ◽  
Specific Staining ◽  
Tetrazolium Chloride ◽  
Tissue Sections ◽  
Blue Tetrazolium Chloride ◽  
In Situ Detection

Using a digoxygenin-labelled DNA probe derived from the porcine repeat element PRE-1, we have developed a protocol for the detection of transplanted porcine islets and hepatocytes against a background of murine host tissue. Analysis of this probe by Southern blotting indicated that PRE-1 hybridizes to pig genomic DNA but not to human or mouse DNA. On tissue sections, hybridizing probe was detected using alkaline phosphatase-conjugated anti-digoxygenin antibody visualized with 5-bromo-4-chloro-3-indolyl-phosphate/4-nitro-blue tetrazolium chloride (BCIP/ NBT) substrate. We have demonstrated sensitive and highly specific staining of porcine nuclei in fixed, paraffin embedded tissue sections, and have applied the technique to detect porcine pancreatic islets and hepatocytes transplanted into murine kidney and spleen. Applications of this technique include detection of transplanted cells or organs across a variety of xenogeneic barriers.

Reactive radical study using the polyvinyl alcohol–potassium iodide solution as a new chemical probe

2021 ◽  
Author(s):  
Hiroto Matsuura ◽  
Nguyen Tran Trung ◽  
Bounyang Ouanthavinsak ◽  
Jin Sakamoto ◽  
Yuichiro Takemura ◽  
...  
Keyword(s):  
Polyvinyl Alcohol ◽  
Potassium Iodide ◽  
Potassium Iodide Solution ◽  
Chemical Probe ◽  
Iodide Solution

The use of potassium iodide solution as photochemical actinometer for vacuum ultraviolet region

2016 ◽  
Vol 38 (2) ◽  
pp. 67-70
Author(s):  
A. V. Mamaenko ◽  
A. O. Samsoni Todorov ◽  
O. V. Zui ◽  
V. A. Yaremenko ◽  
V. V. Goncharuk
Keyword(s):  
Potassium Iodide ◽  
Vacuum Ultraviolet ◽  
Ultraviolet Region ◽  
Potassium Iodide Solution ◽  
Iodide Solution ◽  
Vacuum Ultraviolet Region

Metabolically induced permeability changes across mesothelium and endothelium

1964 ◽  
Vol 206 (2) ◽  
pp. 373-382 ◽  
Author(s):  
Joseph Cascarano ◽  
Arnold D. Rubin ◽  
William L. Chick ◽  
Benjamin W. Zweifach
Keyword(s):  
Carotid Artery ◽  
Tetrazolium Salt ◽  
Phenazine Methosulfate ◽  
Depth Of Penetration ◽  
Tissue Slices ◽  
Tetrazolium Chloride ◽  
Quantitative Studies ◽  
Permeability Changes ◽  
Inner Surface

The ability of mesothelial and endothelial membranes to influence solute passage into tissue preparations was investigated under in vitro conditions. It is possible by using the Warburg formula for the diffusion of solutes into tissue slices to calculate the concentration of solute bathing the subepithelial tissue when the depth of penetration and tissue activity are known. These conditions were met by selecting an electron acceptor with histochemical properties—the tetrazolium salt, 2- p-iodophenyl-3- p-nitrophenyl-5-phenyl tetrazolium chloride (INT). Quantitative studies with hemidiaphragm demonstrate that the concentration of INT which reaches the inner surface of the mesothelial layer is only 9% of that in the medium. Addition of a redox dye, phenazine methosulfate (PM), increases this value by 6.7 times. (Qualitatively similar results were obtained across mesothelial surfaces of liver and heart, as well as at endothelial surfaces of heart and carotid artery.) The ability of PM to increase permeability is counteracted by malate or succinate, and in turn restored when the latter is blocked with malonate. These results suggest that oxidative metabolism and adenosine triphosphate formation are intimately linked with this phenomenon.

Specific Determination of Ethylene Oxide and Ethylene Chlorohydrin in Cosmetics and Polyoxyethylated Surfactants by Gas Chromatography with Electron Capture Detection

1993 ◽  
Vol 76 (2) ◽  
pp. 292-296 ◽  
Author(s):  
Kumiko Sasaki ◽  
Keiji Kijima ◽  
Mitsuharu Takeda ◽  
Shigeo Kojima
Keyword(s):  
Gas Chromatography ◽  
Ethylene Oxide ◽  
Electron Capture ◽  
Potassium Iodide ◽  
Electron Capture Detection ◽  
Potassium Iodide Solution ◽  
Ethylene Chlorohydrin ◽  
Specific Determination ◽  
Iodide Solution ◽  
Nitrogen Stream

Abstract A simple specific determination method was developed for ethylene oxide (EO) and ethylene chlorohydrin (ECH) in cosmetics and surfactants. EO is desorbed from samples by using a nitrogen stream and absorbed into acidic potassium iodide solution, where it is converted to ethylene iodohydrin (EIH). Any remaining ECH in the samples is converted to EO by the addition of sodium hydroxide, and the desorption procedure is repeated with a fresh acidic potassium iodide absorbing solution. EIH is extracted with benzene and determined by gas chromatography with electron capture detection. EO and ECH contents in the samples are calculated from EIH results. Recoveries from water and shampoo samples were 70.3 ± 5.4 and 58.9 ± 1.2%, respectively, for EO and 66.3 ± 4.0 and 64.5 ± 4.6%, respectively, for ECH. Detection limits in 0.2-2.0 g samples were in the 0.005-0.03 μg/g range for EO and 0.01-0.07 μg/g for ECH. High levels of EO (30-394 μg/g as ECH) were found in 5 of 18 polyoxyethylated surfactant samples, but only small amounts (0.07-4.0 μg/g) of ECH were detected in the samples. EO was not detected in cosmetic samples tested, but ECH was present in small quantities (≤1.11 μg/g).

Rapid Single-Step Kinetic Colorimetric Assay for Lactate Dehydrogenase in Serum

1973 ◽  
Vol 19 (2) ◽  
pp. 223-227 ◽  
Author(s):  
Charles C Allain ◽  
Carl P Henson ◽  
M Keith Nadel ◽  
Adam J Knoblesdorff
Keyword(s):  
Lactate Dehydrogenase ◽  
Dehydrogenase Activity ◽  
Dynamic Range ◽  
Colorimetric Assay ◽  
Single Step ◽  
Tetrazolium Salt ◽  
Lactate Dehydrogenase Activity ◽  
Tetrazolium Chloride ◽  
System A ◽  
Reducing Substances

Abstract We report an improved kinetic colorimetric system for measuring lactate dehydrogenase activity in serum. In the system a tetrazolium salt, 2-p-iodophenyl-3-p-nitrophenyl-5-phenyl tetrazolium chloride, is used as the chromogenic indicator of dehydrogenase activity, with diaphorase serving as the electron transfer agent. All ingredients required for an assay are combined in a single dry reagent that is stable at room temperature. The method is 2.5 times as sensitive as the ultraviolet method of Wacker and has a dynamic range three times that of the ultraviolet method. Reducing substances in serum do not affect the results. Precision, range of linearity, and stability of reagent after reconstitution are excellent. Results for fresh sera correlated well with those obtained by the "A-Gent" ultraviolet method (Wacker method at 37°C) and with the SMA 12/60.

Dissolution of bismuth metal by tri-iodide ion in acidified potassium iodide solution

1970 ◽  
Vol 48 (18) ◽  
pp. 2847-2852 ◽  
Author(s):  
Digby D. MacDonald ◽  
G. A. Wright
Keyword(s):  
Metal Surface ◽  
Potassium Iodide ◽  
Potassium Iodide Solution ◽  
Kcal Mole ◽  
Rotating Disc ◽  
Rate Determining Step ◽  
Iodide Ion ◽  
Rate Of Dissolution ◽  
Reaction Proceeds ◽  
Iodide Solution

Dissolution of bismuth metal by tri-iodide ion in acidified potassium iodide medium has been studied using a rotating disc technique. The rate constant for the dissolution process, kT, in solutions 0.5 M in potassium iodide at 25.0 °C has been shown to be related to the square root of the disc angular velocity, ω, by the following expression[Formula: see text]The activation energy for this reaction was found to be 4.7 kcal mole−1. These data clearly establish that the rate of dissolution is determined by the rate at which tri-iodide ion is transported to the metal surface. In solutions of [KI] = 0.10 M, however, the rate determining step appears to involve diffusion of tri-iodide ion through a layer of BiI3 which forms on the bismuth metal surface as the reaction proceeds.

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