34 results about "Thiazole orange" patented technology

The present invention provides methods for determining the presence of immobilized nucleic acid employing unsymmetrical cyanine dyes that are derivatives of thiazole orange, a staining solution and select fluorogenic compounds that are characterized as being essentially non-genotoxic. The methods comprise immobilizing nucleic acid, single or double stranded DNA, RNA or a combination thereof, on a solid or semi solid support, contacting the immobilized nucleic acid with an unsymmetrical cyanine dye compound and then illuminating the immobilized nucleic acid with an appropriate wavelength whereby the presence of the nucleic acid is determined. The cyanine dye compounds are typically present in an aqueous staining solution comprising the dye compound and a tris acetate or tris borate buffer wherein the solution facilitates the contact of the dye compound and the immobilized nucleic acid. Typically the solid or semi-solid support is selected from the group consisting of a polymeric gel, a membrane, an array, a glass bead, a glass slide, and a polymeric microparticle. Preferably, the polymeric gel is agarose or polyacrylamide. The methods employing the non-genotoxic compounds represent an improvement over commonly used methods employing ethidium bromide wherein the present methods retain the advantages of ethidium bromide, ease of use and low cost, but without the disadvantageous, known mutagen requiring special handling and waste procedures.

The invention discloses a method for detecting the activity of DNA methylase and DNA methyltranseferase based on restriction endonuclease Hpa pi and exonuclease III. An unlabeled DNA probe is prepared and the sequence of the probe comprises a cleavage site of the restriction endonuclease and a methylated CpG site. The purpose of detecting the activity of the methylase and methyltranseferase is reached by steps of hybridizing the unlabeled DNA probe with the target DNA, carrying out methylation treatment by the methylase, carrying out the specific cleavage of the restriction endonuclease Hpa pi and enabling exonuclease III having the activity of 3'->5' exonuclease to act on the double-stranded DNA and then adding fluorescence signal molecules thiazole orange which having different signals to single-stranded DNA and double-stranded DNA. According to the method, since no complex material or labeling of DNA probes need to be prepared, the defects of high detection cost, cumbersome operations and poor reproducibility caused by the preparation of the material and labeling of DNA probes are avoided. The method disclosed by the invention has the advantages of low cost and high sensitivity and is fast and simple.

The present invention discloses the solid synthesis process of thiazole orange cyanine dye. The present invention is superior to liquid phase synthesis, and has simple separation and easy purification. Compared with available solid synthesis process, the present invention has the advantages of automatic cutting, simplified synthesis process and lowered production cost. The present invention has solid carrier without volatization, no toxicity, no bad smell and environment friendship.

The invention discloses a method for detecting PARP-1 (Poly ADP-Ribose Polymerase-1) activity based on fluorescent dye TOTO-1 analysis. The method comprises the following steps: (1) mixing and reacting activated DNA, PARP-1 (Poly ADP-Ribose Polymerase-1) and NAD<+> (Nicotinamide Adenine Dinucleotide), and realizing catalyzed synthesis of a PAR polymer (Poly ADP-Ribose) with lots of negative charges by PARP-1; (2) excising double strands from the obtained product by using ExoIII to release PAR; (3) acting TOTO-1 (Thiazole Orange Dimer-1) and the product PAR polymer, and detecting the product solution by utilizing a fluorescence spectrophotometer. The fluorescence signal intensity change is observed by utilizing enhancement of a fluorescence signal generated by combining the TOTO-1 and the product PAR polymer, so the method can be used for detecting the PARP-1. The method disclosed by the invention has the advantages of being simple, convenient, rapid, high in sensitivity and capable ofavoiding from labeling a DNA probe.

The invention belongs to the field of biochemistry and particularly relates to a substituted coumarin-thiazole orange derivative, a preparation method therefor and use of the substituted coumarin-thiazole orange derivative. The substituted coumarin-thiazole orange derivative provided by the invention has a structural formula represented by a formula I shown in the description. The invention also provides the preparation method for the substituted coumarin-thiazole orange derivative and use of the substituted coumarin-thiazole orange derivative in fluorescent recognition of G-quadruplex. The substituted coumarin-thiazole orange derivative provided by the invention has the advantages that the toxicity is low, the selectivity is good, the sensitivity is high, raw materials are simply and easily obtained, the whole synthesis route is high in operability, reaction conditions are relatively mild, the overall cost is relatively low, and the like, thereby undoubtedly having better market competitiveness compared with the existing inefficient G-quadruplex fluorescent probes.

The invention belongs to the field of novel medicines and compounds, and discloses a thiazole orange styrene derivative, a method for preparing the same and application of the thiazole orange styrene derivative to preparing medicines capable of resisting medicine-resistant bacteria. The thiazole orange styrene derivative is of a structure shown as a formula (I). An R in the formula is defined as an instruction book. The method includes carrying out condensation reaction on thiazole orange analogues and different types of aromatic aldehyde to obtain the thiazole orange styrene derivative. The thiazole orange styrene derivative, the method and the application have the advantages that the method is simple, and raw materials for the thiazole orange styrene derivative are easily available; excellent interaction between the thiazole orange styrene derivative and bacterium division proteins FtsZ can be realized, and accordingly reproduction of diversified medicine-resistant bacteria such as vancomycin-resistant enterococci and methicillin-resistant staphylococcus aureus can be inhibited; the thiazole orange styrene derivative is little in toxic and side effect and has a prospect of being developed to obtain novel antibiotic medicines.

The invention discloses PH-sensitive chitosan drug-carrying micelle with targeting and fluorescent characteristics and a preparation method thereof; the multifunctional chitosan drug-carrying micelle with targeting property, PH sensitivity, invisibility and fluorescent tracing combined in a whole is constructed by using PH-sensitive amphiphilic chitosan as drug-carrying micelle and polyethylene glycol as a link arm, and grafting folic acid and fluorescent dye thiazole orange to two ends of the micelle; the chitosan drug-carrying micelle of the invention has good fluorescent properties, and has good drug-carrying property for anticancer drug 5-fluorouracil, with drug-carrying efficiency up to 42.76%; under physiological environmental PH (PH=7.4), accumulative drug release within 8 h is only 2%, accumulative drug release within 26 h is less than 8%, and toxic and side effects of the chitosan drug-carrying micelle for normal tissue cells during transport of the anticancer drug 5-fluorouracil can be decreased; in addition, under tumor cell lysosome PH value (PH=4.5), good PH-sensitive drug release property is exhibited, and 5-fluorouracil release within 4 h is up to 95%.

The invention discloses a method for analyzing and evaluating the erythropoiesis function of mouse marrow through a flow cytometry. According to the method, mouse marrow cells are subjected to dyeing incubation with thiazole orange, rat IgG and APC Rat Anti-mouse Ter119. Incubation sequence is that firstly, the mouse marrow cells and thiazole orange are incubated for appropriate time, centrifugation and supernatant abandoning are conducted, then, rat IgG is added to be incubated for appropriate time, and APC Rat Anti-mouse Ter119 is added to be incubated for appropriate time. Finally, PBS is added, the proportions of mature erythrocytes, reticulocyte, normoblast and proerythroblast in all cells of the mouse marrow are quantitatively analyzed on the flow cytometry, and therefore the erythropoiesis function of the mouse marrow is analyzed and evaluated. The method is quick, accurate, simple, easy to practice, low in cost and beneficial to application and popularization.

The invention discloses a method for detecting phagocyte function based on flow cytometry. In the method of the invention, Escherichia coli bacteria liquid and thiazole orange staining solution are shaken and incubated on a constant temperature shaker for an appropriate time, washed with PBS to remove non-specific adhesion, and centrifuged. Discard the supernatant. Finally, add an appropriate amount of PBS to resuspend, add to the phagocyte culture medium, and incubate in the incubator for an appropriate time. Phagocytes were collected by digestion, and the cell distribution of fluorescent populations was quantitatively analyzed on a flow cytometer, thereby analyzing the function of phagocytes to capture phagocytic bacteria. The invention is fast, accurate, simple and easy to implement, and has low cost, which is beneficial to popularization and application.

The invention discloses a fluorescence detection method of a potassium ion. The specific DNA (5'-GGGTTAGGGTTAGGG TTAGGG-3') of the potassium ion is in an irregular curling state when no potassium ion exists, fluorescent dye thiazole orange and a G-rich sequence are acted to generate very strong fluorescence, after the potassium ion is added, the specific DNA of the potassium ion forms a G tetrahedral structure, and the fluorescence is decreased. The fluorescence detection method disclosed by the invention not only has high sensitivity and better specificity, but also is simple and rapid, and the whole process can be completed in five minutes.

The invention discloses a fluorescent probe, a preparation method thereof and an application of the fluorescent probe for detecting a nucleic acid G-quadruplex structure. The probe is provided with a structure in a general formula (I) is simple and stable in structure and easy to prepare. The probe can be used for specific detection of a nucleic acid G-quadruplex secondary structure, and the G-quadruplex secondary structure in solution can be rapidly detected by a fluorescence spectrometer or direct visual inspection under irradiation of a fluorescent lamp. The probe can be used for detecting the nucleic acid G-quadruplex structure in agarose gel or polyacrylamide gel and can also be used for detecting, marking or displaying existence or distribution of the G-quadruplex structure in living cells. Fluorescent materials have efficient and specific recognition capability for the nucleic acid G-quadruplex structure, have the advantages of excellent cell membrane permeability, low photo-induced toxicity, biological toxicity and photo-bleaching performance and the like, and overcome the shortcomings of high cost, high equipment requirement, complicated technical operation and the like in other detection methods.

The invention provides a carbazole-bridge-based fluorescent cyanine dye probe. The structure of the cyanine dye probe comprises a cyanine dye probe which is formed by respectively bonding carbazole bridge bases between enzoxazol and 4-methyl quinoline salt of thiazole orange and thiazole yellow; one end of a 3-site side chain of a carbazole ring is connected with the 2-C of the enzoxazol, and the other end of the 6-formoxy is connected with 4-vinyl quinoline salt compound. The invention simultaneously provides a preparation method of the carbazole-bridge-based fluorescent cyanine dye probe. According to the invention, the preparation method of the fluorescent dye biological probe is simple, and a raw material is available; by the preparation method, the maximum emission wavelength of the newly synthesized fluorescent dye probe generates red shift, fluorescence strength is enlarged, Stocks displacement is enlarged, fluorescence quantum yield is improved, and light stability is enhanced. In the probe provided by the invention, the advantages of hiazole orange and thiazole yellow embedded fluorescent dye are maintained, thereby being beneficial to further improvement of sensitivity and reduction of dosage as well as improvement of stability. The probe provided by the invention has the characteristics of wide spectral range, large molar extinction coefficient, high capitalization rate, high sensitivity and the like.