A Western Blot assay is performed by performing a probing process on a membrane containing target proteins, by contacting the membrane with a fluorescent resonant energy transfer (FRET) solution and allowing the probing process to proceed for a probing time period. The probing process results in a target protein becoming labeled with both a donor chromophore and an acceptor chromophore, which are effective as a donor-acceptor pair for FRET when so linked to the target protein. While performing the probing process, the labeled target proteins are measured by irradiating the membrane with an excitation light to excite the donor chromophores, wherein in each labeled target protein, the excited donor chromophore transfers energy to the acceptor chromophore by FRET and, in response, the labeled target protein emits an emission light. The intensity of the emission light is then measured. The light measured may be light emitted from the donor chromophore and/or light emitted from the acceptor chromophore.

The present disclosure provides novel serology methods for detecting viremic or latent zoonotic viral infections, such as porcine cytomegalovirus (PCMV) or Porcine lymphotropic herpesvirus (PLHV) infections, in a subject at any stage of development. More specifically, the present disclosure provides an automated multiplexed immunoassay for the rapid and simultaneous detection of one or more anti-PCMV antibodies or anti-PLHV antibodies in biological fluids of a subject suspected of being infected with PCMV or PLHV.

A method for synthesis, assembly and function test of an artificial chloroplast genome of Chlamydomonas reinhardtii. Rational design has been carried out on the Chlamydomonas reinhardtii chloroplast genome for the first time, and total artificial synthesis of the Chlamydomonas reinhardtii chloroplast genome is proposed. By using totally chemically synthesized chloroplast genome segments, total chemical de novo synthesis and assembly of a chloroplast genome are achieved in a yeast-bacterium system. Then, a totally chemically synthesized chloroplast genome is transformed into Chlamydomonas cells to replace the original chloroplast genome, which works normally, and has been verified, fulfilling biological functions of the totally chemically synthesized chloroplast genome. According to the embodiments, the Chlamydomonas reinhardtii chloroplast genome is an efficient platform for carrying out synthetic biology operation.

A protein analysis platform includes a platform body comprising: a gel working unit provided in a top portion of the platform body and comprising a gel accommodation area for accommodating at least one gel; at least one electrophoresis tank provided along a side of the gel accommodation area and provided with at least one electrode; and a blotting layer stack provided in a bottom portion of the gel working unit and comprising an electrode layer; wherein a removable bottom plate is provided between the gel working unit and the blotting layer stack and detachably corresponds to a bottom side of the gel accommodation area. The protein analysis platform is used for western blotting or next-generation western blotting, wherein the protein analysis platform can quickly complete steps such as gel casting, electrophoresis, and blotting in one platform.

An antibody YWHG-1 against HLA-G molecule and use thereof. The antibody is produced by the hybridoma deposited under CCTCC NO: 202120, and is prepared by an immunogen with amino acids which are common to seven known HLA-G isoforms (HLA-G1, HLA-G2, HLA-G3, HLA-G4, HLA-G5, HLA-G6 and HLA-G7) which is located at positions 72-91 (QTDRMNLQTLRGYYNQSEAS) (SEQ ID NO: 19) in the heavy chain al domain of the HLA-G molecule. Nucleotides encoding the antibody YWHG-1 and encoded amino acid sequences thereof are provided as well. The antibody YWHG-1 can be used for immunohistochemistry, immunoblotting, flow cytometry and other assays for HLA-G isoforms.

Disclosed in the present invention are a monoclonal antibody (YWHG-4) against HLA-G isoforms (HLA-G1, HLA-G4, HLA-G5) and use thereof. The antibody (YWHG-4) is produced by the hybridoma deposited under CCTCC NO: 2021204, and an antigenic peptide (RGYYNQSEASSHTLQWMIGC) of amino acid sequences at positions 106-126 of a heavy chain of HLA-G molecules is used as an immunogen. Provided in the present invention are a nucleotide encoding the YWHG-4 antibody of the present invention and an encoded amino acid sequence thereof. Further provided in the present invention is use of the YWHG-4 antibody in the detection of the HLA-G isoforms (HLA-G1, HLA-G4, HLA-G5) by means of immunoblotting, immunohistochemistry, ELISA and flow cytometry.

A method for simulating sensitive response of Jurkat cells in mechanical microenvironments, includes the following steps: S1, transfecting FRET probes into the Jurkat cells by using an electroporation method; S2, simulating the Jurkat cells in the mechanical microenvironments for a first time; S3, collecting FRET images of the Jurkat cells by using a FRET microscope; S4, performing quantitative analysis and statistical analysis on the FRET images; S5, simulating the Jurkat cells in the mechanical microenvironment for a second time; and S6, detecting an ERK phosphorylation level by using a Western blot method. By simulating different states of Jurkat cells, such as no-coated adhesion, charge adsorption, extracellular matrix adsorption, and endothelial cell layer adhesion, and by using methods of FRET living cell observation and biochemical detection method of ERK phosphorylation antibody, the method detects the Jurkat cells in different states, to obtain ERK activity of Jurkat cells in different mechanical microenvironments.

Described herein are detecting methods for conformational disease, aging and proteinopathies, by measuring the presence of b-isox-precipitates and the levels of b-isox-captured proteins in biofluids of healthy individuals and patients. Research identified additional biomarkers, which made it possible to detect, diagnose or treat, a human disease in a human subject by, with or without adding an isoxazole to an obtained biofluid sample, detecting the biomarker. Use of b-iso and/or biomarkers for diagnosing the disease are made possible.

The invention provides a qualitative immunoblot-based in vitro method for the detection of onconeural antibodies class IgG to twelve different antigens (amphiphysin, CV2, PNMa2/Ta, Ri, Yo, Hu, recoverin, SOX1, titin, zic4, GAD65 and Tr (DNER)) in serum or plasma samples of mammalian animals such as dogs, cats, ferrets, and rabbits for early diagnosis of twenty two cancer and cancer-associated neurological diseases. Detection of these antibodies in the blood of the animals is confirmed via an indirect immunofluorescent assay. Examples, including enzyme anti-dog, anti-cat, anti-ferret, and anti-rabbit conjugates, and serum or plasma quantities, are provided.

The present invention relates to a method for selecting lymphocytes recognizing a conformational epitope of a protein antigen from a population of cells, comprising a step of identifying at least one lymphocyte binding to at least two separate peptides comprising sequences of the protein antigen, in particular at least three separate peptides comprising sequences of the protein antigen.