416 results about "Blot" patented technology

The invention describes methods for labeling or detecting of immobilized poly(amino acids), including peptides, polypeptides and proteins, on membranes and other solid supports, using fluorescent dipyrrometheneboron difluoride dyes. Such immobilized poly(amino acids) are labeled or detected on blots or on arrays of poly(amino acids), or are attached to immobilized aptamers.

The invention relates to the technical field of electro-analytical chemistry and protein identification transducers, and discloses a synthesizing method of functionalized ionic liquid and a preparation method and application of a molecular imprinting electrochemical transducer composed of the functionalized ionic liquid, carboxylation multiwalled carbon nanotubes and glassy carbon electrodes. The preparation method of the molecular imprinting electrochemical transducer comprises the steps that the polymerizable amino-functionalized ionic liquid is used as a functional monomer, BSA is used as template protein, N, N'-methylene bisacrylamide is used as a cross-linking agent, an oxidation-reduction system composed of ammonium persulfate and TEMED is used as an initiator, after polymerization, a molecularly imprinted polymer film is formed on the surfaces of the glassy carbon electrodes decorating the carboxylation multiwalled carbon nanotubes, and then the template protein is eluted to obtain the molecular imprinting electrochemical transducer which can identify template protein in a specific mode. The molecular imprinting electrochemical transducer has the advantages of being simple in preparation, low in material cost, high in selectivity, good in biocompatibility, and capable of being used for identifying and detecting protein in aqueous solution.

The invention discloses an up-conversion fluorescence immune chromatography test paper for the quantitative detection of neomycin and a preparation method thereof. The up-conversion fluorescence test paper comprises a support layer, an adsorption layer and a protection layer; the absorbing layer comprises an adsorption fiber layer, a fluorescent antibody fiber layer, a cellulose membrane layer and a water adsorption material layer at a handle hand; the cellulose membrane layer is provided with detection blotting printed by a carrier protein solution coupled with NEO and contrast blotting printed by a goat anti-mouse IgG; the fluorescent antibody adopts an NEO monoclonal antibody or polyclonal antibody marked by NaYF4:Yb:Er nanoparticles. Through the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin, the application of the immune chromatography marked by up-conversion fluorescence nanometer materials in the quantitative detection of NEO residual is realized, so that the detection of the NEO residual is not subjected to background interference; the up-conversion fluorescence immune chromatography test paper for the quantitative detection of the neomycin is strong in specificity, high in sensitivity, simple, intuitional and accurate in detection, low in cost, wide in range of application and easy to popularize and apply.

The invention relates to a Raman spectrum detection method for a molecularly-imprinted membrane (MIM) by use of a Raman spectrometer used as a detection means with the combination of a molecular imprinting technology and a membrane technology and is used for rapidly detecting target molecules-dimethylguanidine hydrochloride and the like in real time. According to the method, a piece of lens paper for an optical lens is used as a substrate (carrier), guanidine hydrochloride or 4-(2-amino ethyl) benzsulfamide is used as an imprinted template, nanogold / silver sol is added in a pre-polymerization solution, and a polymer is directly light-gathered on the lens paper, so as to be convenient to carry and detect. The addition of the nanogold / silver sol is similar to the surface enhanced raman spectroscopy in principle, the response strength of the Raman spectrum can be enhanced and the fluorescence signal interference is reduced; the molecularly-imprinted membrane after and before target adsorption is compared with a standard substance, so that the peaked change in the Raman spectrum can be observed and whether a practical sample contains a target molecule can be judged.

The invention discloses an immune chromatography test paper strip for the quantitative determination of clenbuterol based on up-conversion fluorescent nanoparticle label and a preparation method thereof. The test paper strip comprises a supporting layer, an adsorption layer, and a protection layer, wherein the adsorption layer comprises an adsorption fibrous layer, a fluorescent antibody fibrous layer, a cellulose film and an absorbent material layer at the handle end, the cellulose film is provided with detection blots printed by using a CL coupling carrier protein solution and control blots printed by using goat anti-mouse IgG antibodies, and the fluorescent antibody is an NaYF4:Yb:Er nanoparticle labelled CL monoclonal antibody or polyclonal antibody. According to the invention, the application of immune chromatography based on the up-conversion fluorescent nanoparticle label in the quantitative determination of CL residues is realized, so that the detection of the CL residues has no background interference; and the test paper strip has the advantages of strong specificity, high sensitivity, simple, and accurate detection, low cost, wide application scope, and easiness in popularization and application.

The invention relates to a test strip for rapidly detecting residues and a preparation method of the test strip. The bottom layer of the test strip is used as a supporting layer; an intermediate adsorption layer is fixed on the supporting layer; an external protection layer is fixed on the adsorption layer; the adsorption layer comprises an adsorption fiber layer, a gold-labeled antibody fiber layer for adsorbing and coupling lincomycin and a cellulose membrane layer sequentially arranged on the testing sample end and further comprises a water absorbing material layer arranged on the handle end; a detection blot printed by a lincomycin carried protein solution and a control blot printed by a goat anti- or rabbit anti-mouse IgG antibody solution are arranged on the cellulose membrane layer. The test strip is high in detection specificity, high in sensitivity and simple and fast in operation; detection results are displayed visually, directly and accurately; a special apparatus and a professional are not in need in the detection; and the test strip has the following characteristics of low cost, low investment as well as easiness for promotion and application.

The present invention provides a gene encoding a recombinant porcine circovirus type 2 Cap protein and application thereof. The present invention aims to obtain a soluble recombinant PCV2 Cap protein, and adopts a PCR method for fusion of MPG and NLS partially deleted Cap protein N-terminal gene to construct a new gene; and then the new gene is cloned into an E. coli expression vector pET28a to obtain a recombinant plasmid; the recombinant plasmid is transformed into E. coli BL21 to obtain recombinant engineering bacteria; the recombinant engineering bacteria is subjected to induced expression by IPTG to obtain the soluble recombinant PCV2 Cap protein. SDS-PAGE and Western-blot identification shows that most of the recombinant protein MrCap is present in the bacterial lysis supernatant and is soluble; and the recombinant protein MrCap has high immunogenicity and immunoreactivity, and lays foundation for the development of a PCV2 antibody detection kit and subunit vaccines.

The present invention includes a method for detecting and isolating sugarcane proteins that interact with the HC-Pro and P1 proteins of SrMV and other proteins involved in gene silencing, particularly in sugarcane. The method uses a two hybrid assay with an HC-Pro, P1, or other silencing-related protein-containing bait protein and a prey protein containing a polypeptide encoded by a DNA molecule in a cDNA library. The method also includes identification of false positives through reverse two-hybrid assays and using in vitro techniques such as farwestern blots or pull down assays where plant physiological conditions may be replicated. Finally, interactions may be confirmed in planta. Some novel proteins used in and discovered using the these methods are also identified. Methods of using viral and plant proteins to regulate silencing in plants such as sugarcane are also discussed.

A preparation method of core-shell molecular imprinting nanometer silicon spheres comprises the following steps: synthesizing double bond-modified nanometer silicon spheres; synthesizing an auxiliary identification polymer chain with an identification group and an anchoring group which are randomly distributed; and self-assembling immunoglobulin heavy-chain binding protein BiP with the auxiliary identification polymer chain to prepare double-bond modified nanometer silicon spheres, and carrying out surface polymerization to form an imprinted polymer layer and an elution template. According to the invention, the prepared core-shell structured imprinting nanometer silicon spheres make imprinted sites be positioned on the material surface, substantially increase the imprinting specific surface area, and greatly increase the adsorption efficiency and adsorption quantity to target proteins; the existence of the auxiliary identification polymer chain makes imprinting have a specificity and a high efficiency, and the combination of the specificity and the high efficiency realizes the high-expansion and high-efficiency enrichment of natural microproteins and a strong practicality; and the use of cloned BiP as a template in the invention overcomes a problem that the natural microproteins cannot be obtained.

Many of the effects of nitric oxide are mediated by the direct modification of cysteine residues resulting in an adduct called a nitrosothiol. A method to detect proteins which contain nitrosothiols involves several steps. Nitrosylated cysteines are converted to tagged cysteines. Tagged proteins can then be detected, for example, by immunoblotting and / or can be purified by affinity chromatography. The method is applicable to the detection of S-nitrosylated proteins in cell lysates following in vitro S-nitrosylation, as well as to the detection of endogenous S-nitrosothiols in selected protein substrates.