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Chicken PEBP1 gene polyclonal antibody, preparation method and application thereof

A polyclonal antibody and gene technology, applied in the biological field, can solve problems such as blanks, and achieve the effects of strong specificity, wide application range, and good sensitivity

Pending Publication Date: 2021-06-11
JIANGXI AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the research on chicken PEBP1 gene-related antibodies is still blank

Method used

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  • Chicken PEBP1 gene polyclonal antibody, preparation method and application thereof
  • Chicken PEBP1 gene polyclonal antibody, preparation method and application thereof
  • Chicken PEBP1 gene polyclonal antibody, preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 The preparation method of broiler chicken PEBP1 gene polyclonal antibody

[0030] 1. Construction of PET32a-PEBP1 recombinant expression vector

[0031] 1.1 According to the chicken-derived PEBP1 gene nucleotide sequence reported in Genbank, high-quality nucleotide coding sequences were screened by bioinformatics analysis, and then specific primers were designed and synthesized, and the corresponding combination with the template was performed to amplify the PEBP1 gene and PEBP1 gene. The total PCR reaction system was 20 μl, see Table 1.

[0032] Table 1 General system of PCR reaction

[0033]

[0034] PCR reaction program: pre-denaturation at 94°C for 5 min, denaturation at 94°C for 30s, annealing at 56°C for 35s, extension at 72°C for 40s, 35 cycles, and finally extension at 72°C for 10 min.

[0035] Depend on figure 2 A It is easy to know that based on the chicken PEBP1 gene sequence published in GenBank, a pair of primers with strong specificity ...

Embodiment 2

[0062] Example 2 Establishment of an indirect immunofluorescence detection kit for chicken PEBP1 protein

[0063] Antigen preparation: Samples were collected from five related tissues (pulmonary artery, heart, lung, liver and kidney) from broilers, ducks, mice, rabbits and goats and stored in a 4% paraformaldehyde solution for later use Perform the corresponding section preparation.

[0064] Blocking: After antigen retrieval, dewaxed glass slides with a thickness of about 5 μm were blocked in goat serum for 30 min.

[0065] Sample to be tested and detection antibody: Incubate the slides with the anti-PEBP1 protein antibody (1:200 dilution) and pre-immune serum (1:200 dilution) prepared in Example 1 at 4°C overnight;

[0066] Washing: wash 3 times with PBS-T (5min each time);

[0067] Primary and secondary antibodies: Incubate slides with 1:300 diluted Cy3 conjugated Goat Anti-Rabbit IgG (H+L) incubation solution (Boster, Wuhan, China) for 50 min at room temperature in the da...

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Abstract

The invention discloses a chicken PEBP1 gene polyclonal antibody, a preparation method and application thereof, and belongs to the technical field of biology. The preparation method comprises the step of immunizing New Zealand white rabbits by taking PEBP1 recombinant protein as an antigen to obtain the polyclonal antibody. The method can be used for preparing the high-quality polyclonal antibody, and the polyclonal antibody can be used for detecting the PEBP1 protein in animal bodies, has the advantages of wide application range, high specificity, simplicity and convenience in operation, lower cost and the like, and also lays a material foundation and technical support for research and diagnosis of pathological mechanisms of various broiler diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, and relates to a chicken PEBP1 gene polyclonal antibody, a preparation method and application, in particular to a chicken PEBP1 gene polyclonal antibody related to the occurrence and development of broiler pulmonary hypertension and ascites diseases, a preparation method and an application. Background technique [0002] Broiler ascites syndrome (AS) was once called poultry edema, congestive heart failure and pulmonary hypertension syndrome, etc. It is a relatively common metabolic disorder and non-infectious disease. The disease was first discovered in high-altitude and cold areas, and its main features are increased cardiac load, hypoxemia, right heart hypertrophy, etc. After pathological autopsy, it can be seen that a large amount of light yellow transparent fluid accumulated in the abdominal cavity, pericardial effusion, kidney and The liver is congested and enlarged or even necrotic. Studies hav...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K16/06C12N15/70C12N15/12A61K39/395A61P9/12A61P7/10A61P9/04G01N33/68
CPCC07K16/18C07K16/06C12N15/70C07K14/465A61P9/12A61P7/10A61P9/04G01N33/6893A61K2039/505A61K2039/552G01N2333/465G01N2800/321G01N2800/325
Inventor 郭小权胡国良施燕刘平顾玥明张彩英刘佩李庆庆郭凤萍程素芳
Owner JIANGXI AGRICULTURAL UNIVERSITY
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