214 results about "Western blot" patented technology

The invention discloses an anti-CD3D antibody and application thereof.The antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of CDR-H1 of the heavy chain variable region is shown as SEQ ID NO: 4, the amino acid sequence of CDR-H2 of the heavy chain variable region is shown as SEQ ID NO: 5, and the amino acid sequence of CDR-H3 of the heavy chain variable region is GDL; the amino acid sequence of the CDR-L1 of the light chain variable region is as shown in SEQ ID NO: 6, the amino acid sequence of the CDR-L2 of the light chain variable region is as shown in SEQ ID NO: 7, and the amino acid sequence of the CDR-L3 of the light chain variable region is as shown in SEQ ID NO: 8. The anti-CD3D antibody is a rabbit monoclonal antibody, the obtained antibody is high in titer, clear in specific positioning in immunohistochemical staining and free of non-specific background staining, the antibody is wide in application, and the antibody can be used in pathological immunohistochemical staining experiments and can also be used in Western blot and flow cytometry experiments.

The invention belongs to the technical field of image processing, and discloses a western blot image generation forgery detection method and system, a terminal and a storage medium, and the method comprises the steps: obtaining a to-be-detected image; wherein the to-be-detected image comprises a face image and a western blot image; performing feature extraction on the to-be-detected image based on a counterfeit detection model to obtain visual features; performing generation discrimination on the visual features to obtain a real discrimination result or a counterfeit discrimination result of the to-be-detected image; source discrimination is carried out on the visual features corresponding to the counterfeit discrimination result, and a discrimination result that the to-be-detected image comes from the face image or the western blot image is obtained; and outputting the western blot image to generate a forgery detection result. According to the method, the forgery recognition precision of the western blot image is improved, and the generalization ability of image forgery detection is enhanced.

The invention relates to an application of a THBS4 inhibitor in preparation of a medicine for preventing and / or treating pulmonary arterial hypertension. Three pulmonary arterial hypertension rat models (a hypoxia induction model, a hypoxia combined SUGEN induction model and a monocrotaline induction model) are constructed, the expression change of THBS4 is verified through qRT-PCR and western blot, and the correlation between the expression level of THBS4 and pulmonary arterial hypertension is verified in combination with in-vitro cell experiments and in-vivo intervention. Results show that THBS4 is remarkably up-regulated in pulmonary arteries of the three models, the expression level of THBS4 is positively correlated with the severity of diseases, and THBS4 is an important regulatory factor for promoting vascular remodeling in pulmonary arterial hypertension. In vivo, by knocking down THBS4, pulmonary vascular remodeling and right ventricular hypertrophy are remarkably relieved, and it is prompted that THBS4 inhibitors show good application prospects in the aspect of preventing and / or treating pulmonary arterial hypertension.

The invention discloses a construction and verification method of an HEK293T cell line capable of stably overexpressing human ANT2, which comprises the following steps: cloning a human ANT2 gene, constructing the human ANT2 gene into a lentivirus expression plasmid pLVX-TRE3G, co-transfecting the lentivirus expression plasmid pLVX-TRE3G and a helper plasmid pLVX-Tet3G into an HEK293T-ACE2 cell, and carrying out puromycin and G418 pressurized screening and Western blot identification to obtain the HEK293T cell line capable of stably overexpressing human ANT2. A result shows that the expression quantity of the ANT2 is the highest when the HEK293T-ACE2-ANT2 cell is induced for 12 hours at the Doxycycline of 6mg / L; when the ANT2 is induced to express, the virus nucleic acid load and the N protein expression level after SARS-CoV-2 infection are obviously reduced, which indicates that the ANT2 has an inhibition effect on SARS-CoV-2 infection replication, and a cell model and an experimental basis are provided for exploring the effect of the ANT2 in virus infection resistance.

The invention provides an immunofluorescence detection method and system for dynamically detecting mouse uterus structure development based on multiple time points, and immunofluorescence staining comprises the steps of sheet unfolding, dewaxing, repairing, cell membrane punching, sealing, primary antibody treatment and secondary antibody treatment. The system comprises a sample pretreatment control module, a multi-channel confocal microscope imaging module and a data analysis module integrating fluorescence intensity calculation, Pearson co-localization analysis and a Western blot data integration algorithm. Through a multi-time-point and multi-index immunofluorescence detection method, dynamic and systematic data support is provided for mouse uterus development mechanism research, and an application basis is provided for fundamental research of reproductive biology and construction of related disease models.

The invention relates to the technical field of biological medicines, and discloses a blocking antibody targeting CXCL6 and an application of the blocking antibody in preparation of antitumor drugs. The antibody or antigen-binding fragment thereof is capable of binding specifically to human CXCL6 protein with high affinity (e.g., Kd < = 1 nM). According to the invention, the specificity and high affinity of the antibody are verified by Western Blot, immunofluorescence, immunohistochemistry and surface plasmon resonance (SPR) technologies. Functional experiments show that the antibody can effectively block CXCL6-induced neutrophil chemotaxis, including in Transwell, a three-dimensional gel model and a zebra fish living body model. Besides, in a mouse MC38 colon cancer model, the antibody shows remarkable anti-tumor activity and can inhibit tumor growth driven by CXCL6. The invention also provides a pharmaceutical composition containing the antibody, and application of the antibody in preparation of drugs for preventing or treating diseases (such as inflammatory diseases, autoimmune diseases and cancers) related to abnormal expression or activity of CXCL6.

The present application relates to the technical field of biotechnology, in particular to a programmed death receptor ligand 2 monoclonal antibody and application thereof. The present application provides a PD-L2 monoclonal antibody, amino acid sequences of three CDR regions of a heavy chain thereof; amino acid sequences of three CDR regions of a light chain thereof; or amino acid sequences obtained by substitution, deletion or addition of one or more amino acids of the amino acid sequences, or amino acid sequences functionally identical or similar to the amino acid sequences; or amino acid sequences having at least 80% homology with the sequences. The present application widens the possibility of antibody therapy for blocking PD1 / PD1 ligand, and compared with existing PD-L2 monoclonal antibodies, the 8E3 antibody has unique amino acid sequences and CDR region sequences, high affinity and high specificity. The 8E3 antibody can be used in ELISA, Western blot and flow cytometry detection, and has wide application.

The invention discloses application of a PRTFDC1 gene in promoting porcine skeletal muscle satellite cell differentiation, myotube formation and skeletal muscle development. According to the invention, overexpression and knock-down experiments of the PRTFDC1 are respectively carried out in pig MuSCs, and immunofluorescence, qRT-PCR and Western blot analysis are carried out on cells subjected to induced differentiation, so that the PRTFDC1 is effectively shown to play a positive regulation role in a pig MuSCs differentiation process, and a new marker gene which can be applied to pig molecular breeding and growth performance evaluation is provided; and a novel molecular target and a novel detection tool are provided for muscle development regulation and biological breeding.

The invention relates to the field of antibodies, and particularly provides a high-affinity PADI2 antibody and a preparation method thereof. According to the invention, eight high-affinity PADI2 antibodies are obtained by screening through an antibody pair screening experiment, and the antibodies can be specifically combined with PADI2 protein and have high affinity. Experimental results show that the antibody disclosed by the invention shows excellent binding capacity in ELISA (Enzyme-Linked Immunosorbent Assay), Western blot and cell experiments, and can be used for detecting, diagnosing and treating PADI2 related diseases.

The present application relates to the field of biotechnology and in vitro diagnosis, and relates to a monoclonal antibody of eosinophil peroxidase and application thereof. The antibody of the present application is mainly used for detecting the content of eosinophil peroxidase by western blot. The monoclonal antibody of eosinophil peroxidase of the present application is prepared by immunizing Balb / c mice with recombinant eosinophil peroxidase as an immunogen, then fusing the spleen with myeloma cells NS1, and screening and purifying. The prepared monoclonal antibody of eosinophil peroxidase specifically recognizes the linear epitope of eosinophil peroxidase.

The invention discloses an application of a compound fresh bamboo juice in a guinea pig asthma model, and relates to the field of medicines, a Hartley albino guinea pig is adopted to construct an OVA induced asthma model, and the model is verified to be successfully constructed through evaluation indexes; evaluating the efficacy of the successfully constructed guinea pig asthma model by using different doses of compound fresh bamboo juice, and evaluating the efficacy of the guinea pig asthma model by scoring guinea pig capsaicin stimulation cough inducing times, alveolar lavage fluid leukocyte number, inflammatory factor content, lung tissue hydroxyproline content and behavioral signs. And evaluating the treatment effect of the compound fresh bamboo juice through Western blot detection, lung histopathology and immunofluorescence staining indexes. In a guinea pig asthma model, high-dose and medium-dose groups of the compound fresh bamboo juice can be used for remarkably reducing the number of cough, reducing the number of white blood cells in alveolar lavage fluid and the level of inflammatory factors and inhibiting pulmonary fibrosis, and a good treatment effect is shown. The results provide powerful support for potential application of the compound fresh bamboo juice in asthma treatment.

The invention provides an application of 2-indolecarboxylic acid in preparation of a tumor targeted therapy drug. Through computer-aided drug design and molecular docking optimization, 2-indolecarboxylic acid shows high binding energy to an HER2 kinase structural domain. In HER2 positive NCI-N87 gastric cancer cells, the 2-indolecarboxylic acid significantly inhibits cell proliferation and induces G0 / G1 phase arrest and apoptosis. The mechanism research shows that the 2-indoleformic acid can play a role by regulating and controlling the HER2 / Akt / beta-catenin pathway (Western blot verification). In an NCI-N87 cell tumor-bearing nude mouse model, 2-indolecarboxylic acid (15 mg / kg) significantly inhibits tumor growth, and does not cause obvious weight loss or organ toxicity. Immunohistochemical analysis shows that the positive rate of a tumor tissue proliferation marker Ki-67 in a treatment group is remarkably reduced, and apoptosis detection shows that the proportion of TUNEL positive cells is remarkably increased, which indicates that 2-indolecarboxylic acid inhibits tumor cell proliferation and promotes apoptosis at the same time. The invention provides a lead compound with clinical transformation potential for HER2 targeted therapy.

The invention relates to a high-affinity mouse anti-carp IgM (Immunoglobulin M) monoclonal antibody, a hybridoma cell strain and application of the high-affinity mouse anti-carp IgM monoclonal antibody and the hybridoma cell strain. The mouse anti-carp IgM monoclonal antibody is secreted by the hybridoma cell strain CcM-12A12G11C9C7 with the preservation number of CCTCC (China Center For Type Culture Collection) NO: C2024236. The mouse anti-carp IgM monoclonal antibody can be used for Western blot and ELISA specific detection of IgM in a carp secretion form, and can be used for flow cytometry identification of carp IgM + B cells. The monoclonal antibody is good in specificity and high in sensitivity, can be used for structural analysis, immune response level detection and the like of carp IgM, and lays a foundation for in-depth study of a carp immune system and establishment of a vaccine immune effect evaluation method taking the antibody level as an index.

The invention discloses a construction method of a cabbage efficient genetic transformation regeneration system. The method comprises the following steps: (1) culturing aseptic seedlings; (2) preparing an explant; (3) preparing agrobacterium dip dyeing liquid carrying a target carrier; (4) carrying out dip dyeing and co-culture on the explants; (5) callus differentiation and bud induction; and (6) growing seedlings and rooting, and carrying out PCR (Polymerase Chain Reaction) and Western blot identification on the obtained plants. By optimizing key parameters and vector selection of the cabbage genetic transformation method, compared with the prior art, the method has the advantages that antibiotics are not needed for preliminary screening in the regeneration process, hormone is not needed in the rooting process, the cost is low, the transformation period is short (greater than or equal to 45 days), the regeneration frequency is high, the transformation efficiency is high, and subsequent related protein research work can be conveniently carried out; the important value is realized on shortening the cabbage gene function research period.

The invention discloses application of ailanone in preparation of a medicine for treating feline infectious peritonitis, and belongs to the field of biological medicine. The change of the expression quantity of the feline infectious peritonitis virus N protein is detected through indirect immunofluorescence and western blot, and the change of the quantity titer of the feline infectious peritonitis virus is detected through TCID50. Results show that ailanone has an obvious effect of resisting the feline infectious peritonitis virus, the inhibitory effect on the feline infectious peritonitis virus is enhanced along with increase of the drug concentration, and the half effective concentration is 0.09 mu M. Meanwhile, drug inhibition tests show that ailanone has no inhibition effect on feline calicivirus. The invention provides a novel therapeutic drug and strategy for treating feline infectious peritonitis.

The invention discloses a down-regulated CDC20-based breast cancer cell ferroptosis sensitivity detection method, and belongs to the technical field of detection methods, and the method comprises the following steps: cell grouping and transfection: culturing a breast cancer cell system, and setting an experimental group and a transfection negative control group; wherein in the experimental group, CDC20-siRNA transfection is carried out on breast cancer cells; an experimental group with the CDC20 relative expression quantity lower than 0.5 is screened through Western blot to serve as an optimal experimental group, and subsequent experiments are conducted; and then ferroptosis induction and ferroptosis induction and inhibition are carried out, and related detection is carried out. According to the invention, CDC20 is used as a regulation target of breast cancer ferroptosis sensitivity for the first time, and the target limitation of a traditional method is broken through; and the reliability and repeatability of a detection result are ensured through a standardized operation process and closed-loop verification logic.

The invention relates to the technical field of genetic engineering, in particular to a small yellow croaker IRE1 alpha protein polyclonal antibody as well as a preparation method and application thereof. The invention provides a preparation method of a small yellow croaker IRE1alpha protein polyclonal antibody. The preparation method comprises the following steps: cloning an open reading frame of a small yellow croaker IRE1alpha gene, and constructing a small yellow croaker IRE1alpha gene recombinant expression vector pET-30a (+)-IRE1alpha. Performing inducible expression and purification on the recombinant protein of the Larimichthys polyactis IRE1 alpha, and performing specific analysis on Western Blot; and preparing the polyclonal antibody of the small yellow croaker IRE1 alpha. According to the invention, a recombinant expression vector of the small yellow croaker IRE1alpha gene is constructed by adopting molecular biology and genetic engineering methods, purified recombinant protein is obtained through induced expression and affinity chromatography, the prepared polyclonal antibody can be used for expression detection of the small yellow croaker IRE1alpha protein, and an important material foundation is laid for research on functions and action mechanisms of the small yellow croaker IRE1alpha protein.

The invention belongs to the field of pharmacy, and relates to a protein degradation targeting chimera (PROTAC) taking PD-L1 as a target spot and medical application thereof. The protein degradation targeting chimera is a compound composed of a target protein ligand, an E3 ligase ligand and a linker and a pharmaceutically acceptable salt of the compound, the specific structure of the compound is as shown in a formula I, and substituent groups are described in the specification in detail. A Western blot experiment (Western Blot) shows that the compound disclosed by the invention can be used for degrading the PD-L1 protein in a targeted manner.

The invention discloses an effect and mechanism research method of GOT1 in lung adenocarcinoma cells, and relates to the technical field of biomedical research. Comprising the following steps: S1, cell culture and protein extraction: culturing Bas-2b, A549, H1573 and Calu-3 cells, cleaning the cells with PBS, performing pyrolysis treatment, collecting supernate as a total protein solution, and determining the protein concentration per tube by BCA; s2, carrying out Western Blot experiment, taking 10 micrograms of protein as a loading amount, carrying out electrophoresis, transferring a film, sealing, incubating a primary antibody and a secondary antibody, exposing and developing, and storing a picture; the experiment is repeated for multiple times, and Image J is used for analyzing the strip exposure diagram. According to the method disclosed by the invention, effective knock-down of GOT1 in A549 and H1573 cells is successfully realized through the steps of screening proper cell screening concentration by puromycin, exploring an MOI value, transfecting lentivirus, screening puromycin and the like; the invention provides an important tool and method for follow-up research on the function of GOT1 in lung adenocarcinoma cells.

The invention provides application of lncPRPD regulation and control on GLRX5 in improving pork quality, the lncPRPD is lncRNA obtained by performing whole transcriptome sequencing analysis on longissimus dorsi muscle of Laiwu pigs, and the nucleotide sequence of the lncPRPD is as shown in SEQ ID NO: 1. The method comprises the following steps: firstly, synthesizing a small interfering RNA (Ribonucleic Acid) of the lncPRPD / GLRX5 and constructing a gene overexpression vector of the lncPRPD / GLRX5; then, detecting the combination of the lncPRPD and the GLRX5 protein by utilizing a Pulldown experiment and a Western blot experiment; the expression quantity of the GLRX5 is detected after interference and overexpression of the lncPRPD, and it is found that the lncPRPD can promote expression of the GLRX5 protein level; through detection experiments such as qPCR (quantitative polymerase chain reaction), Western blot, oil red O staining and the like, the lncPRPD and the GLRX5 are detected to be capable of promoting the differentiation of the pig precursor fat cells.

The invention discloses a method for improving autism behaviors through Npas4 expression and overexpression in a cat chat syndrome model, and belongs to the technical field of animal model construction. Selecting SD wild rats and CdCS model rats, and normally feeding and breeding the rats; the method comprises the following steps: measuring the protein concentration by adopting a BCA method, then carrying out Western Blot, then carrying out brain stereotactic injection, and finally carrying out behavioral testing. According to the invention, a CdCS rat model is established by using a CRISPR-Cas9 technology, and the model shows cognitive and social behavior disorders and medial prefrontal cortex (mPFC) neuron dendritic abnormality; in addition, an abnormally expressed gene is found in the model mouse, the research focuses on Npas4, and a new insight is possibly provided for disease treatment.

The invention belongs to the technical field of bioengineering, and particularly relates to a preparation method and application of an N protein polyclonal antibody of a horse coronavirus. The polyclonal antibody is prepared by truncation expression of the horse coronavirus N protein, and a material is provided for research on inhibition of host innate immune mechanism by the horse coronavirus N protein. Prokaryotic expression is carried out on the N protein of the equine coronavirus, the polyclonal antibody is prepared, and a method and a material are provided for molecular research and mechanism research of the equine coronavirus. According to the invention, in combination with bioinformatics analysis of an N protein of the horse coronavirus, a pGEX-6p-N3 recombinant plasmid is constructed, and prokaryotic expression is carried out, so that a purified recombinant protein is obtained. After a mouse is immunized by the recombinant protein, a polyclonal antibody is obtained, an indirect ELISA method detects that the polyclonal antibody has good titer, Western Blot detection has good reactivity, and the polyclonal antibody can be applied to an indirect immunofluorescence test.

This invention belongs to the field of traditional Chinese medicine treatment of acute kidney injury, and provides a traditional Chinese medicine composition for treating sepsis-related acute kidney injury and its preparation method. It is prepared by mixing honeysuckle, forsythia, roasted peach kernel, raw rhubarb, raw astragalus, wine-processed angelica, cinnamon twig, mirabilite, and raw licorice in a certain proportion. It detoxifies and removes blood stasis, invigorates qi and unblocks the meridians. It is used for persistent high fever and difficulty in urination and defecation caused by internal accumulation of toxic heat and blood stasis. Symptoms include oliguria or dribbling, red tongue, and rapid pulse; it is also used for sepsis patients with the above symptoms. Treatment with the detoxifying and blood-stasis-removing granules improved survival rate, effectively restored some renal function, reduced renal tubular damage, and decreased TNF-α and IL-1β expression. Immunohistochemical and Western blot results were consistent. The detoxifying and blood-stasis-removing granules can alleviate sepsis-induced acute kidney injury.

The utility model discloses Western blot full-automatic multi-round circulating detection equipment, which realizes continuous detection of multi-target proteins on a single transfer printing film by integrating a liquid treatment module, an incubation reaction module, a signal capture module and a program control module. Residues are removed by adopting antibody denaturation liquid, signal crosstalk is avoided by combining a chemiluminescence system, 20 cycles of cyclic operation are supported, and a self-cleaning function is achieved. Equipment is compatible with a conventional transfer printing film, and the detection flux and the standardization level are remarkably improved.

The invention provides an application and a method of an RSAD2 agonist glycopyrronium in resisting RSV infection. The method comprises the following steps: S1, virtual screening and molecular docking; s1.1, performing virtual screening and molecular docking; s1.2, carrying out molecular dynamics simulation; s2, cell culture and RSV infection; s3, performing RT-qPCR detection on the expression of the RSAD2 and the virus load of the RSV; s4, performing Westernblot analysis on the influence of a small molecule compound on the expression of the RSAD2; s5, CCK8 is used for detecting the toxicity of the compound to Beaas-2b cells; s6, carrying out indirect immunofluorescence detection on a fluorescence signal of the RSV; s7, animal model establishment; s8, carrying out HE dyeing; s9, carrying out enzyme-linked immunosorbent assay (ELISA); and S10, statistical analysis. According to the present invention, the small molecule agonist targeting the host immune factor RSAD2 is screened, the endogenous antiviral factor is activated from the host perspective, the drug resistance problem caused by direct virus targeting is avoided, the RSAD2 is adopted as the host natural antiviral molecule, the activation strategy has good targeting property and good safety, and the efficient anti-RSV effect under the low toxic-side effect is achieved.

The invention discloses a preparation method of brassinolide derived carbon quantum dots and application of the brassinolide derived carbon quantum dots in plant hormone receptor detection. According to the method, BRs is used as a precursor, CQDs with a BRs semi-reserved structure are prepared through co-carbonization with citric acid, and the nano material with the particle size of 3.1 + / -0.5 nm and the surface rich in hydroxyl / keto is obtained after dialysis and purification. When the CQDs are combined with the BRs receptors, the fluorescence intensity of the CQDs is linearly enhanced along with the concentration of the receptors, the nano sensor constructed based on the CQDs can realize rapid quantitative detection of the BRs receptors in rice leaves, the linear range is 0.01-4.0 mg / mL, the detection limit is as low as 0.003 mg / mL, and the sensitivity is improved by 10 times compared with that of ELISA (Enzyme-Linked Immunosorbent Assay). By detecting the receptor expression quantity of a wild type and BRs receptor overexpression strain in a germination period, a heading period and a mature period, it is verified that the result of the strain is highly consistent with that of Western Blot, and the detection time is shortened to 30 minutes. The method does not need targeted modification, has the advantages of high specificity, interference resistance and low cost, and provides an efficient detection tool for rice molecular breeding and stress resistance regulation.

This application applies to the field of genetic engineering technology and provides an mlaC knockout mutant strain, its construction method, and application. The mlaC knockout mutant strain is obtained by knocking out the mlaC gene in highly virulent Klebsiella pneumoniae (hvKP). The examples of this application innovatively discovered that hvKP lacking the mlaC gene exhibited reduced lung growth and lung damage in mice, as well as reduced mortality in mice, indicating that the MlaC protein is a key pathogenic factor in hvKP. Furthermore, secretory protein profiling and Western blot results showed that the MlaC protein can be secreted extracellularly and is present in all Klebsiella pneumoniae (KP) strains. Its sequence is conserved, making it a potential new vaccine target for KP. This application broadens research on the pathogenic mechanism of KP and provides a theoretical basis for its prevention and treatment.

The invention belongs to the technical field of biological genes, and discloses a method for identifying RNA interaction protein based on a Turbo ID technology and an MCP / MS2 system, the method can be used for identifying protein interacting with RNA in vitro or in vivo, and the method specifically comprises the following steps: firstly, respectively constructing RNA expression vectors inserted with MS2 tags, then constructing fusion protein vectors expressing MCP-Turbo ID, and finally constructing fusion protein vectors for expressing MCP-Turbo ID; the method comprises the following steps of: inserting RNA (Ribonucleic Acid) into an MS2 tag, transgenosis into a plant receptor by utilizing an agrobacterium tumefaciens mediated genetic transformation method to obtain a transgenic plant material expressing MCP-Turbo ID, combining the RNA inserted with the MS2 tag with the MCP-Turbo ID in vivo / in vitro by utilizing a hybridization or in-vitro incubation method, further enabling the Turbo ID to be adjacent to the RNA, biotinylating the RNA binding protein, and obtaining a transgenic plant material expressing the MCP-Turbo ID. And then enriching biotinylated protein by adopting streptavidin magnetic beads, and identifying and analyzing through Western Blot and mass spectrometry. The method not only solves the problem that the interaction protein of specific RNA in a living cell environment cannot be effectively identified in the prior art, but also can capture a transient or low-affinity RNA-protein interaction relationship.

The invention discloses a CD147 nano antibody D2 as well as a preparation method and application thereof, and belongs to the technical field of molecular biology and immunology. The preparation method comprises the following steps: screening from an alpaca nano antibody phage library by utilizing a phage display technology, and sequencing to obtain 9 CD147 nano antibodies; 3 nano antibody genes are selected and cloned into pCold II, and prokaryotic expression recombinant plasmids are successfully constructed; the method comprises the following steps: carrying out inducible expression for 20 hours under the conditions that the final concentration of IPTG (isopropyl-beta-d-thiogalactoside) is 0.2 mmoL / L, the temperature is 16 DEG C and the speed is 100 rpm / min, carrying out nickel column purification, and then carrying out Coomassie brilliant blue dyeing and Western blot identification to determine and obtain the CD147 nano antibody which is named as C36, D2 and H4; the binding activity of the three nano antibodies and the CD147 protein is detected through indirect ELISA (enzyme-linked immuno sorbent assay). The result shows that the CD147 antigen and the three nano antibodies have binding activity and are dose-dependent. Under the same conditions, when the incubation concentration of the nano-antibody is as low as 1.25 micrograms / mL, the D2 binding effect is the best, the amino acid sequence of the nano-antibody is shown as SEQ ID NO.8, and the nucleotide sequence of the nano-antibody is shown as SEQ ID NO.10. The invention provides a more accurate method for detecting the CD147.