CN113801232A - 123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit - Google Patents
123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit Download PDFInfo
- Publication number
- CN113801232A CN113801232A CN202010532117.XA CN202010532117A CN113801232A CN 113801232 A CN113801232 A CN 113801232A CN 202010532117 A CN202010532117 A CN 202010532117A CN 113801232 A CN113801232 A CN 113801232A
- Authority
- CN
- China
- Prior art keywords
- rasal2
- antibody
- phosphorylation
- protein
- antigen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/40—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against enzymes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/44—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material not provided for elsewhere, e.g. haptens, metals, DNA, RNA, amino acids
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/90—Enzymes; Proenzymes
- G01N2333/914—Hydrolases (3)
Abstract
The invention belongs to the technical field of biology, and particularly relates to a 123 th phosphorylation antibody of Rasal2, a preparation method thereof, an ELISA detection kit for detecting 123 th phosphorylation of Rasal2 and a detection method. The invention comprises the following steps: synthesizing an antigenic peptide phosphorylated at the S123 site; coupling the antigen peptide with the hemocyanin to obtain an immune antigen, then immunizing an animal by using the antigen and collecting antiserum; identifying and purifying antiserum to obtain 123 th phosphorylation specific antibody of Rasal 2; an ELISA detection kit for detecting 123 th phosphorylation of Rasal2 protein is obtained by coating an ELISA plate with 123 th phosphorylation antibody of Rasal2 and assisting other detection materials. The method can be used for identifying the 123 th phosphorylation content of the Rasal2 protein in the tissue sample, and has the advantages of simple and convenient operation, strong specificity, high sensitivity and easy standardization.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to preparation of a 123 th phosphorylation antibody of Rasal2 and an ELISA (enzyme-Linked immuno sorbent assay) detection kit for detecting 123 th phosphorylation of Rasal 2.
Background
The prior art discloses Rasal2 as one member of RasGAP family, which inhibits the activity of Ras mainly by promoting the hydrolysis of Ras-GTP to GDP form, thus inhibiting the occurrence and development of tumors in most tumors. Research shows that Rasal2 has two different action modes of promoting and inhibiting tumor growth in breast cancer, and Rasal2 has low expression in luminal B breast cancer and inhibits tumor growth and invasion by inhibiting Ras activity. However, in triple negative breast cancer Rasal2 is overexpressed and, by interacting with ARHGAP24, activates RAC1 to promote invasion and migration of cells.
Studies have disclosed Ras overactivation as an important factor in tumor development and progression. Rasal2 is considered to be a tumor suppressor, which is expressed in a low proportion of cancer tissues, such as luminal B breast cancer, lung cancer, ovarian cancer, bladder cancer, nasopharyngeal cancer and the like, however, in Triple Negative Breast Cancer (TNBC), Rasal2 is overexpressed, which simultaneously promotes tumor invasion and carcinogenesis, and in colorectal cancer, Rasal2 can promote the occurrence and development of colorectal cancer by up-regulating LATS2/YAP1 signaling pathway, and thus, the role of Rasal2 in cancer is related to different types of tumors and different subtypes of the same tumor.
To date, there has been no phosphorylated Rasal2 detection antibody that is useful as a detection marker for the development of tumors. Based on the basis and the current situation of the prior art, the inventor of the application intends to provide a 123 th phosphorylation specific antibody of Rasal2 protein, a preparation method and an ELISA detection kit.
Disclosure of Invention
The invention aims to provide a specific antibody for identifying the phosphorylation site of Rasal2 protein S123 and a preparation method thereof based on the foundation and the current situation of the prior art.
A further object of the present invention is to provide an ELISA detection kit for detecting 123 th phosphorylation of Rasal 2.
It was found that even though Rasal2 promoted or inhibited two diametrically opposite effects in breast cancer, Rasal2 phosphorylation at position 123 was universal and consistent in breast cancer. After the site is phosphorylated, autophagy can be effectively induced, cells are protected to reduce apoptosis induced by external stress, and proliferation of tumor cells is promoted, and clinical data show that 123 th phosphorylation of Rasal2 is obviously negatively related to prognosis and 5-year survival period of a breast cancer patient; research results show that 123 th phosphorylation of Rasal2 is likely to be used as a biomarker of breast cancer, and a certain reference basis is provided for drug resistance research and accurate treatment of the breast cancer, so that the simple, rapid and high-sensitivity detection of 123 th phosphorylation level of Rasal2 in cells or tissues has important significance for disease diagnosis, targeted drug screening and the like.
More specifically, the present invention is to provide a novel,
the invention provides an antibody, which recognizes 123 th phosphorylation of Rasal2 protein.
The antibody is prepared by the following method:
immunizing animals by using antigenic peptide containing 123 th phosphorylation of Rasal2 protein and collecting antiserum;
the 123 th phosphorylation antibody of Rasal2 was isolated from the antiserum.
The antigenic peptide contains the following sequence: EQQTDSTKGRC (SEQ ID NO 1); when the antigenic peptide is produced, cysteine (Cys) may be added to the end or the front end of the antigenic peptide to obtain the antigenic peptide.
In a preferred embodiment of the present invention, S in EQQTDSTKGRC is a phosphorylation site, and a C is added before E to obtain the antigenic peptide phosphorylated at the S123 site (i.e., phosphorylated at 123 th site of Rasal2 protein) shown in fig. 1.
In another aspect, the invention provides methods for producing the antibodies.
The preparation method of the Rasal2 protein phosphorylation Ser123 site-specific antibody is carried out according to the following steps: firstly, synthesizing 123 th phosphorylated amino acid sequence of Rasal 2; coupling the synthesized antigen peptide with the hemocyanin to obtain a complete antigen, then immunizing a rabbit by using the antigen, and collecting antiserum; and thirdly, purifying and firming the obtained antiserum to finally obtain the Rasal2 phosphorylated S123 site-specific antibody.
For example, in a preferred embodiment of the invention, the antibody preparation method comprises the steps of:
(1) synthesizing 123 th phosphorylated antigen peptide of Rasal2 protein;
(2) coupling the 123 th phosphorylated antigen peptide of the Rasal2 protein with the hemocyanin to obtain an immune antigen, then immunizing an animal by using the antigen and collecting antiserum;
(3) separating and purifying the antiserum obtained in the step (2) to obtain the 123 th phosphorylation specific antibody of Rasal 2.
The invention also provides application of the antibody, namely recognition of 123 th phosphorylation of Rasal2 protein.
Preferably, the antibody can be used for preparing a reagent for disease diagnosis or screening of targeted drugs.
Correspondingly, the invention provides a detection kit, which contains an antibody for recognizing 123 th site phosphorylation of Rasal2 protein.
Preferably, the detection kit comprises an ELISA plate coated with a Rasal2 protein phosphoric S123 acidified antibody and/or a phosphate buffer solution, a confining liquid, a substrate buffer solution and a stop solution TMB reaction liquid.
The ELISA detection kit for detecting 123 th phosphorylation of Rasal2 specifically comprises the following components:
an enzyme label plate coated with a p-Rasal2(Ser123) phosphorylation antibody;
p-Rasal2(Ser123) phosphorylating antibody;
③ the washing buffer solution is PBST which is 1000mL of 0.01mol/L PBS +0.5m L Tween-20;
fourthly, the blocking solution is TBST with 3 percent BSA;
coating buffer solution: 0.015M Na2CO3, 0.035M NaHCO3, pH 9.6;
sixthly, TMB mother liquor (2mg/m L stock solution): 10mg of TMB was dissolved in 5m L absolute ethanol sufficiently,
substrate reaction solution: 0.5m L TMB mother liquor, 10m L substrate buffer solution and 2.1 mu L H2O2(30 percent) are prepared fresh when in use;
the stop solution is 2M H2SO 4.
Preferably, the detection kit also contains a Rasal2 protein S123 phosphorylation antibody.
The kit can be used for identifying S123 phosphorylation of Rasal2 protein, for example, Rasal2 protein phosphorylated at 123 th site or peptide fragment thereof is detected by using an Elisa method.
Preferably, the ELISA plate in the detection kit of the invention is coated with the 123 th phosphorylation antibody of Rasal2, and correspondingly, the preparation method of the detection kit further comprises the step of coating the 123 th phosphorylation antibody of Rasal2 on the ELISA plate.
The invention has the beneficial effects that:
A. through the identification of the prior phosphoprotein mass spectrometry, the 123 th serine in the Rasal2 in the breast cancer tissue is found to be phosphorylated under the sugar-free condition or the anti-cancer drug treatment condition, the antibody is prepared by taking the 123 th serine phosphorylation site and the upstream and downstream conserved amino acid sequences of the Rasal2 protein as antigen peptides, coupling the antigen with the haemocyanin to obtain a complete antigen, immunizing a New Zealand rabbit, and purifying.
B. The preparation method of the 123 th phosphorylation form antibody of Rasal2 provided by the invention is helpful for revealing other functions besides RasGAP activity of Rasal2 protein, provides the 123 th phosphorylation of Rasal2 as an important basis of a biomar of breast cancer, and provides a certain reference for drug resistance research and accurate treatment of the breast cancer.
C. The ELISA detection kit for detecting 123 th phosphorylation of Rasal2 is formed by using the p-Rasal2 antibody to coat an ELISA plate and other ELISA conventional reagents, expression change conditions of p-Rasal2 of breast cancer cells under different stress conditions are detected by using the kit, and data results indicate that a lot of antitumor drugs inhibit tumor generation by improving the activity of AMPK, and the level of p-Rasal2 is probably improved to promote the generation of drug resistance of the tumor.
D. The p-Rasal2 antibody provided by the invention has high purity and good specific binding capacity.
E. The ELISA detection kit for detecting the 123 th phosphorylation of Rasal2 provided by the invention is simple in operation, high-efficiency, sensitive and easy to standardize.
Drawings
FIG. 1 is a schematic representation of the Ser123 phosphorylation site obtained in the first embodiment;
FIG. 2 shows the result of Western blotting detection specific to the anti-p-Rasal 2(Ser123) antibody;
FIG. 3 shows the expression of p-Rasal2 in breast cancer cells after ELISA method to detect different conditions.
Detailed Description
The present invention is further illustrated by the following examples, wherein the materials and equipment used in the present invention are conventional in the art unless otherwise specified.
EXAMPLE 1 preparation of Ser123 phosphorylated Rasal2 complete antigen
Firstly, designing an antigen peptide phosphorylated at an S123 site, as shown in figure 1, and simultaneously adding cysteine (Cys) at the front end or the tail end according to the amino acid characteristics of a polypeptide sequence so that the antigen peptide can be coupled with the hemocyanin;
preparing a hemocyanin solution, and dissolving the hemocyanin in PBS to ensure that the final concentration is 2 mg/ml;
mixing the hemocyanin solution with the Sulfo-SMCC solution, uniformly mixing at room temperature and incubating for 4 hours;
dialyzing by using PBS to remove free SMCC, and calculating the protein concentration by using a broadford method;
fifthly, mixing the synthesized antigen peptide phosphorylated at the S123 locus with the hemocyanin-SMCC solution, uniformly mixing at room temperature and incubating for 4 hours to prepare the 123 th phosphorylation antigen of Rasal 2;
example 2 preparation of Rasal2 phosphorylated antibody
Adding equivalent Freund complete adjuvant into 123 th phosphorylation complete antigen of Rasal2 and emulsifying;
secondly, the emulsified antigen is injected into the backs of New Zealand rabbits at multiple points in the skin, and the dosage of each rabbit is 0.5mg/kg (basic immunity);
③ after 20 days of basic immunization, the antigen is fully mixed and emulsified with equal volume of Freund incomplete adjuvant;
fourthly, the emulsified antigen is injected into the back of the New Zealand rabbit at multiple points in the skin for the first boosting immunity;
after 20 days, performing secondary boosting immunization according to the operation steps in the fourth step;
sixthly, after 14 days of the second boosting, the immune animals are sacrificed and blood is taken. Standing the serum in ice water bath for 4h, centrifuging for 10min at 5000 rpm, and taking out the supernatant to obtain antiserum;
seventhly, purifying the p-Rasal2(Ser123) antibody by affinity chromatography, and using a phosphorylated synthetic peptide coupled with a hemocyanin and agarose binding material as a packing material of a chromatographic column.
Example 3 specific identification of the p-Rasal2(Ser123) antibody
Constructing a Rasal 2S 123A mutant and Myc-Rasal2-S123A, respectively transferring recombinant vectors Myc-Rasal2-S123A and Myc-Rasal2-WT into 293T cells, then adding glucose starvation treatment for 4 hours, and performing co-immunoprecipitation through M2-Beads, thereby identifying whether the p-Rasal2 antibody can recognize two differently expressed proteins, and determining the specificity of the antibody;
the results are shown in FIG. 2, where WT: 293T cell transfected with Myc-Rasal2-WT expression vector, S123A: 293T cells of point-transfer mutation Myc-Rasal 2-S123A;
the experimental result shows that the obtained anti-p-Rasal 2(Ser123) antibody can specifically recognize 123 th phosphorylation of Rasal2, and the phosphorylation of Rasal2(Ser123) can be obviously increased by glucose starvation.
Example 4 ELISA detection kit for detecting 123 rd phosphorylation of Rasal2
The detection kit comprises the following components:
an enzyme label plate coated with a p-Rasal2(Ser123) phosphorylation antibody;
p-Rasal2(Ser123) phosphorylating antibody;
③ the washing buffer solution is PBST which is 1000mL of 0.01mol/L PBS +0.5mL of Tween-20;
fourthly, the blocking solution is TBST with 3 percent BSA;
coating buffer solution: 0.015M Na2CO3, 0.035M NaHCO3, pH 9.6;
sixthly, TMB mother liquor (2mg/mL stock solution): 10mg of TMB was dissolved well in 5mL of absolute ethanol,
substrate reaction solution: 0.5mL of TMB mother liquor, 10mL of substrate buffer solution and 2.1 mu L H2O2(30 percent), and is prepared fresh when in use;
the stop solution is 2M H2SO 4.
Example 5 detection of phosphorylation of Rasal2 using ELISA detection kit for 123 rd phosphorylation of Rasal2
The method comprises the following steps:
coating antigen: uniformly mixing 100ul of Rasal2 123 th site phosphorylation antibody and 900ul of coating buffer solution, adding into an enzyme label plate, and coating overnight at 4 ℃;
washing the coated enzyme label plate for 6 times, and patting the enzyme label plate on gauze;
adding 100 ul/hole of a sample to be tested, incubating for 1.5 hours in a constant temperature box at 37 ℃, and then washing for 3 times;
adding 100ul of sealing liquid into each hole, incubating for 1.5 hours in a constant temperature box at 37 ℃, and then washing for 3 times;
adding primary antibody, diluting according to a ratio of 1:10000, adding 100ul of primary antibody into each hole, simultaneously keeping negative control and blank control, incubating for 1.5 hours in a constant temperature box at 37 ℃, and then washing for 3 times;
sixthly, adding secondary antibody, diluting according to a ratio of 1:10000, adding 100ul of secondary antibody into each hole, simultaneously keeping a negative control and a blank control, incubating for 1.5 hours in a constant temperature cabinet at 37 ℃, and then washing for 3 times;
seventhly, adding TMB substrate color development liquid, incubating for 30min at the temperature of 37 ℃ in a dark place at 50 ul/hole;
adding stop solution into the mixture, 50ul per hole, and detecting the OD450 value of each hole in a microplate reader.
FIG. 3 shows that the agonists of AMPK can effectively promote phosphorylation at 123 th position of Rasal2 through the detection result of an ELISA kit under different conditions.
The above description is only for the specific embodiments of the present application, but the scope of the present application is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present disclosure should be covered within the scope of the present application. Therefore, the protection scope of the present application shall be subject to the protection scope of the claims.
Sequence listing
<110> university of Compound Dan
<120> Rasal2 protein 123 th phosphorylation specific antibody, preparation method and ELISA detection kit
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 11
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Glu Gln Gln Thr Asp Ser Thr Lys Gly Arg Cys
1 5 10
Claims (10)
1. An antibody which recognizes phosphorylation 123 of Rasal2 protein.
2. The antibody of claim 1, prepared by the following method:
immunizing animals by using antigenic peptide containing 123 th phosphorylation of Rasal2 protein and collecting antiserum;
the 123 th phosphorylation antibody of Rasal2 was isolated from the antiserum.
3. The antibody of claim 2, wherein said antigenic peptide has the sequence shown in SEQ ID NO 1.
4. A method for producing the antibody of claim 1, which comprises the steps of:
(1) synthesizing 123 th phosphorylated antigen peptide of Rasal2 protein;
(2) coupling the 123 th phosphorylated antigen peptide of the Rasal2 protein with the hemocyanin to obtain an immune antigen, then immunizing an animal by using the antigen and collecting antiserum;
(3) separating and purifying the antiserum obtained in the step (2) to prepare a 123 th phosphorylation specific antibody of Rasal 2.
5. Use of the antibody of claim 1 in the preparation of a preparation that recognizes phosphorylated Rasal2 at position 123.
6. The use according to claim 5, wherein the antibody is used for the preparation of a reagent for the diagnosis of a disease or for the screening of targeted drugs.
7. A detection kit is characterized by comprising an antibody for recognizing 123 th site phosphorylation of Rasal2 protein.
8. The detection kit as claimed in claim 7, wherein the detection kit comprises an ELISA plate coated with an acidified antibody against Rasal2 protein phosphate S123 and/or a phosphate buffer solution, a blocking solution, a substrate buffer solution and a stop solution TMB reaction solution.
9. The test kit of claim 7, further comprising Rasal2 protein S123 phosphorylation antibody.
10. Use of the test kit according to claim 7, characterized in that the 123 th phosphorylated Rasal2 protein is detected using the Elisa method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010532117.XA CN113801232A (en) | 2020-06-11 | 2020-06-11 | 123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010532117.XA CN113801232A (en) | 2020-06-11 | 2020-06-11 | 123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113801232A true CN113801232A (en) | 2021-12-17 |
Family
ID=78943927
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010532117.XA Pending CN113801232A (en) | 2020-06-11 | 2020-06-11 | 123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113801232A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116444677A (en) * | 2023-06-09 | 2023-07-18 | 中国人民解放军军事科学院军事医学研究院 | FoxM1 protein Y575 phosphorylated polyclonal antibody, preparation method and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090081659A1 (en) * | 2007-03-07 | 2009-03-26 | Cell Signaling Technology, Inc. | Reagents for the detection of protein phosphorylation in carcinoma signaling pathways |
| US20110059463A1 (en) * | 2009-07-09 | 2011-03-10 | Cell Signaling Technology, Inc. | Serine and Threonine Phosphorylation Sites |
-
2020
- 2020-06-11 CN CN202010532117.XA patent/CN113801232A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090081659A1 (en) * | 2007-03-07 | 2009-03-26 | Cell Signaling Technology, Inc. | Reagents for the detection of protein phosphorylation in carcinoma signaling pathways |
| US20110059463A1 (en) * | 2009-07-09 | 2011-03-10 | Cell Signaling Technology, Inc. | Serine and Threonine Phosphorylation Sites |
Non-Patent Citations (1)
| Title |
|---|
| XUAN WANG等: "Phosphorylated Rasal2 facilitates breast cancer progression", 《EBIOMEDICINE》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116444677A (en) * | 2023-06-09 | 2023-07-18 | 中国人民解放军军事科学院军事医学研究院 | FoxM1 protein Y575 phosphorylated polyclonal antibody, preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1727896B (en) | Phosphorylation and corresponding non-phosphorylating protein polyclonal antibody are preparing application in reagent for disease diagnosis and preparation method thereof | |
| KR20130102688A (en) | Multiple biomarker set for breast cancer diagnosis, method of detecting the same, and diagnosis kit for breast cancer using antibody against the same | |
| CA2783308C (en) | Method for diagnosing malignant tumor | |
| CN114181308B (en) | Procalcitonin antibody and application thereof | |
| CN104991077A (en) | Troponin I competition turbidimetry detecting kit | |
| Asano et al. | Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay | |
| CN113801232A (en) | 123 th phosphorylation specific antibody of Rasal2 protein, preparation method and ELISA detection kit | |
| CN102680706B (en) | Application of protein CTSF (Cathepsin F) in preparation of reagent for diagnosing gastric cancer and diagnostic reagent kit | |
| KR20150041620A (en) | Pivka-ⅱ measurement method, measurement reagent, and measurement kit | |
| WO2011146479A1 (en) | Method and composition for the diagnosis and monitoring of inflammatory diseases | |
| CN105158474A (en) | Reagent kit for detecting serum HER2 and application | |
| AU2018323281A1 (en) | Methods for detecting cancers, and detection reagent | |
| US8975030B2 (en) | Method for cancer detection | |
| CN113429470A (en) | Amino acid sequence of Survivin epitope for detecting esophageal cancer marker and application thereof | |
| KR20180115922A (en) | TPM4-ALK fusion protein and composition for diagnosing cancer | |
| US4731336A (en) | Immunoassay for complement fragments | |
| CN101796415B (en) | Novel liver cancer marker | |
| KR101919403B1 (en) | Recombinant protein and use thereof | |
| JP2007071849A (en) | Diagnostic agent and diagnostic kit for mesothelioma | |
| KR101225846B1 (en) | Assay method for human orotate phosphoribosyltransferase protein | |
| CN109810184A (en) | People NF155 antigen, people's NF155 antibody assay kit and the preparation method and application thereof | |
| Nagamine et al. | Development of a high sensitivity ELISA for the assay of metallothionein | |
| CN109061163A (en) | A kind of CST1 chemiluminescence detection kit and its detection method | |
| KR20180115923A (en) | hnRNPA2B1-FAM96A fusion protein and composition for diagnosing cancer | |
| CN102135542A (en) | ELISA (Enzyme Linked Immunosorbent Assay) method and kit for detecting soluble protein PDCD5 (Programmed Cell Death 5) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20211217 |
|
| WD01 | Invention patent application deemed withdrawn after publication |