Detection kit for blood serum PSMD4 protein, detection method for blood serum PSMD4 protein and application of detection kit

A detection kit and the technology of the kit, applied in the fields of immunology and biology, can solve the problems such as the application of the ELISA detection kit for PSMD4 protein that has not been reported in the literature

Active Publication Date: 2017-06-23
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, there is no literature report on the accurate detection of PSMD4 protein in serum samples of tumor patients; there is no accurate, simple and sensitive detection method for PSMD4 protein content in serum samples of tumor patients; there is no literature report on the detection of PSMD4 protein. ELISA detection kit and the application of such kit in tumor serological diagnosis and screening

Method used

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  • Detection kit for blood serum PSMD4 protein, detection method for blood serum PSMD4 protein and application of detection kit
  • Detection kit for blood serum PSMD4 protein, detection method for blood serum PSMD4 protein and application of detection kit
  • Detection kit for blood serum PSMD4 protein, detection method for blood serum PSMD4 protein and application of detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Preparation of detection ELISA plate: Coating buffer 1×PBS, pH: 9.6 Dilute the antibody (rabbit anti-human PSMD4 polyclonal antibody) to 2.0ug / ml, add 100 μl to each well of the ELISA plate, After sealing the plate, incubate overnight at 4°C for later use.

[0063]Detection of serum samples and recombinant proteins: Shake off the liquid in each well, add ELISA microplate washing solution (1×PBS solution containing 0.05% Tween-20), 300 μl per well, wash 2-3 times, and dry; Block with a blocking solution containing 3% bovine serum albumin in PBS solution, add a sample at a volume of 200ul / well, and block for 2 hours at room temperature: shake off the liquid in each well, add ELISA plate washing solution, 300μl per well , washed 2-3 times, dried, serum samples were diluted 1:1 with sample diluent 1×PBS (pH:7.4), added at a volume of 100 μl / well, incubated at room temperature for 2 hours; PSMD4 recombinant protein was diluted with samples Dilute the solution into different...

Embodiment 2

[0064] Example 2: Optimization

[0065] 1. Optimization of coating antibody concentration:

[0066] Select different coating antibodies rabbit anti-human PSMD4 polyclonal antibody (PTG company and Abnova company), choose different concentrations (7.5μg / ml, 5μg / ml, 2.0μg / ml, 1μg / ml, 0.5μg / ml, 0.25 μg / ml) coated ELISA plate, detected according to the operating steps in Example 1, adding known PSMD4 protein standards of different concentrations. According to the obtained OD value, select the blank group with the smallest OD value and the closest linear relationship between the OD value and the standard protein concentration as the best antibody (PTG company). The coating antibody concentration is 2.0 μg / ml.

[0067] 2. Optimization of blocking solution:

[0068] Choose different blocking solutions, containing 1% bovine serum albumin + PBS solution, containing 2% bovine serum albumin + PBS solution, containing 3% bovine serum albumin + PBS solution, containing 4% bovine serum al...

Embodiment 3

[0075] Embodiment 3: ELISA kit detects PSMD4 fusion protein

[0076] Dilute the PSMD4 recombinant protein (PTG#ag6691) doubly, starting from 10ng / ml, serially dilute to 7 concentrations, 10ng / ml, 3ng / ml, 1ng / ml, 0.3ng / ml, 0.1ng / ml, 0.03ng / ml, 0.01ng / ml each sample set up 2 duplicate wells, 100 μl / hole, repeat the detection 5 times according to the step of embodiment 1, the result is similar, as figure 1 As shown, the reaction between the PSMD4 recombinant protein and the antibody has a good concentration-dependent relationship and an obvious linear relationship.

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Abstract

The invention belongs to the technical field of immunology and biology and particularly relates to a detection kit for a blood serum PSMD4 protein and a detection method for the blood serum PSMD4 protein and application of the detection kit. The ELISA detection kit and detection method for the PSMD4 protein, provided by the invention, can be applied to the serological diagnosis and tumor screening of a variety of malignant tumors, particularly lung cancer, gastric cancer and colon cancer. According to the detection kit, the detection method and the application, the operation is simple and convenient, the content of the PSMD4 protein in blood serum of malignant tumor patients can be detected in an accurate and high-sensitivity manner, and thus, a new means and a method are provided for clinical checking and fundamental researches.

Description

technical field [0001] The invention belongs to the field of immunology and biotechnology, in particular to an ELISA detection kit for PSMD4 protein and its detection method and its application in serological diagnosis and tumor screening of various malignant tumors (lung cancer, gastric cancer and colorectal cancer) . Background technique [0002] Cancer refers to all malignant tumors. According to the statistics of the World Health Organization, there are more than 14 million new cancer patients in the world every year, and China has become the country most seriously affected by cancer in the world. The incidence of cancer in mainland China accounts for almost half of the global cancer incidence. Cancer ranks second among various causes of death in China, and ranks first in cities. Among them, the mortality rate of lung cancer has risen the fastest, ranking first in the mortality rate of malignant tumors. Followed by liver cancer, gastric cancer and colorectal cancer. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/574G01N33/543
CPCG01N33/543G01N33/574G01N33/577G01N33/68G01N33/5748G01N33/57488
Inventor 丁劲李晓峰程卓
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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