CN111363031A - pSer131 polyclonal antibody of BNIP3, preparation method and application thereof - Google Patents
pSer131 polyclonal antibody of BNIP3, preparation method and application thereof Download PDFInfo
- Publication number
- CN111363031A CN111363031A CN202010140528.4A CN202010140528A CN111363031A CN 111363031 A CN111363031 A CN 111363031A CN 202010140528 A CN202010140528 A CN 202010140528A CN 111363031 A CN111363031 A CN 111363031A
- Authority
- CN
- China
- Prior art keywords
- bnip3
- antibody
- protein
- phosphorylation
- phosphorylated
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/82—Translation products from oncogenes
-
- 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
- C07K16/065—Purification, fragmentation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- 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
- G01N33/574—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
- G01N2333/82—Translation products from oncogenes
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Oncology (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Hospice & Palliative Care (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a BNIP3 protein Ser131 site phosphorylation antibody and a preparation method and application thereof. The amino acid sequence of the antigen synthetic peptide is SSHCD (S-p) PPRSQ, and the phosphorylation site is serine at the 131 th site of BNIP3 protein. The BNIP3 protein S131 site phosphorylation antibody can not only recognize exogenously overexpressed phosphorylated BNIP3, but also recognize endogenously phosphorylated BNIP3 of cells, and can also be applied to BNIP3 protein stability, mitochondrion autophagy and apoptosis determination.
Description
Technical Field
The invention belongs to the technical field of biomedicine for preparing antibodies, and particularly relates to a BNIP3 protein Ser131 site phosphorylation antibody and a preparation method and application thereof.
Background
BNIP3 is a mitochondrial outer membrane protein, belongs to BCL-2 family, and early researches find that BNIP3 plays an important role in regulating apoptosis, and more evidence shows that BNIP3 can be used as a receptor mediated mitochondrion autophagy in recent years. In rat primary cultured cardiomyocytes and various tumor cell lines, it was observed that BNIP3 could be modified by phosphorylation, however the phosphorylation site and corresponding function were not answered. Zhu et al found that phosphorylation of Ser17 and Ser24 sites of BNIP3 had an important role in mediating mitochondrial autophagy by BNIP3 through site mutation in 2013, but kinases directed to the sites have not been found so far, indicating that phosphorylation of the two sites is not necessarily true. Therefore, it is necessary to further study the new phosphorylation sites and develop a BNIP3 protein phosphorylation antibody to explore the regulatory role of BNIP3 in mediating mitophagy or apoptosis.
On the other hand, BNIP3 was observed to be rapidly degraded in earlier studies, and BNIP3 was later found to be degraded by the ubiquitin-proteasome pathway. Generally, ubiquitination of proteins is closely related to phosphorylation, but how BNIP3 phosphorylation has a regulatory role in its ubiquitination degradation is not clear. Therefore, it is necessary to develop a BNIP3 protein phosphorylation antibody to ascertain the precise regulation of BNIP3 phosphorylation on its ubiquitination degradation. At present, the domestic and foreign markets lack of the BNIP3 antibody which can be used for researching endogenous phosphorylation, so that related research and application cannot be carried out.
Disclosure of Invention
The invention aims to provide a polyclonal antibody phosphorylated by serine at 131 th position of BNIP3, a preparation method and application thereof.
An antigen synthetic peptide of a BNIP3 protein phosphorylation antibody, wherein the amino acid sequence of the antigen synthetic peptide is SSHCD (S-p) PPRSQ, and the phosphorylation site is serine at the 131 th position of BNIP3 protein.
A preparation method of BNIP3 protein phosphorylation antibody comprises the following steps:
1) coupling the antigen synthetic peptide with carrier protein to obtain antigen;
2) immunizing an animal by using the antigen obtained in the step 1), and collecting antiserum;
3) and (3) carrying out affinity column separation and purification on the antiserum obtained in the step 2) by using the synthesized modified peptide to obtain a Ser131 site phosphorylation antibody of the BNIP3 protein.
Preferably, the immunization mode in the step 2) is specifically as follows: 1 priming injection and 4 booster injections; the priming and boosting injections were both: the antigen synthetic peptide and adjuvant are combined to immunize animals, and the antigen synthetic peptide and adjuvant are injected subcutaneously on two sides of a part with thin and loose neck and back skin, the two sides of gluteus and thighs are respectively injected intramuscularly, the two sides of waist are injected intradermally, and the claw pad is injected.
Preferably, in the step 3), the purification is performed by: determining the titer of the antiserum before purification to the antigen by Dot blot experiment (Dot blot), and determining whether the antiserum before purification can specifically identify the antigen; and purifying the collected antiserum by using an affinity separation-affinity purification cycle technology to obtain the polyclonal antibody phosphorylated by the serine at the 131 th position of BNIP 3.
Use of the BNIP3 protein phosphorylation antibody described above for dot blot detection.
The use of the BNIP3 protein phosphorylation antibody described above for immunoblot detection.
Use of the BNIP3 protein phosphorylating antibody described above for detecting exogenous overexpression of phosphorylated BNIP3 in a cell.
Use of the BNIP3 protein phosphorylating antibody described above for detecting endogenously expressed phosphorylated BNIP3 in cells.
The invention has the beneficial effects that: the technical scheme of the invention lists the BNIP3 protein Ser131 site phosphorylation antibody, the antigen synthetic peptide thereof and the preparation method thereof, wherein the BNIP3 protein Ser131 site phosphorylation antibody can detect the phosphorylation level of BNIP3 protein, can be used for evaluating the effect of BNIP3 mediated mitochondrion autophagy or apoptosis and the regulation and control effect of BNIP3 phosphorylation on ubiquitination degradation thereof, thus being helpful to research the effect of BNIP3 protein phosphorylation in a cell signal path related to BNIP3 and a tumorigenesis mechanism, and providing a potential effect target spot for the diagnosis or treatment of clinical tumor diseases; meanwhile, the BNIP3 protein Ser131 site phosphorylation antibody can be directly applied to tumor diagnosis, treatment and prognosis judgment. The antibody can be used for detection of Dot Blot (Dot Blot), immunoblot (Western Blot) and the like. This antibody, in addition to recognizing exogenously overexpressed phosphorylated BNIP3, also recognized cellular endogenously phosphorylated BNIP 3. The present inventors successfully developed a polyclonal antibody phosphorylated on serine 131 of BNIP3 against phosphorylated BNIP 3.
Drawings
FIG. 1 shows the result of observing the change in bands using Western blot after mutating different positions of BNIP3 in example 1 of the present invention.
FIG. 2 is a graph of Dot blot with pre-purified antiserum as the primary antibody in example 3 of the present invention.
FIG. 3 is a graph of Dotblot using a polyclonal antibody phosphorylated with serine 131 of BNIP3 as a primary antibody in example 4 of the present invention.
FIG. 4 is a Western blot chart of the polyclonal antibody with phosphorylated serine at 131 th position of BNIP3 as a primary antibody in example 4 of the present invention.
FIG. 5 is phosphorylated BNIP3 detected exogenously using polyclonal antibodies phosphorylated on serine 131 at BNIP3 as primary antibodies.
FIG. 6 shows the detection of endogenously phosphorylated BNIP3 in cells using polyclonal antibody phosphorylated on serine 131 at BNIP3 as a primary antibody.
Detailed Description
In order that the invention may be more fully understood, reference will now be made to the following description. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete.
Example 1 determination of the phosphorylation site of BNIP3 protein
The potential phosphorylation sites in the BNIP3 amino acid sequence were first screened for possible phosphorylation sites in the BNIP3 protein by the bioinformatics software Netphorest 2.0 and Netphos 2.0, and then determined by mass spectrometry literature and search of the protein phosphorylation site database PhosphoSiteplus. Then, the positions are mutated one by one, and the change of the BNIP3 protein band is observed by using an immunoblot (Western blot), so that the specific phosphorylation site of the BNIP3 protein is finally determined to be the 131 th amino acid-serine Ser131, and the result is shown in FIG. 1 (7 th lane).
Example 2 Synthesis of antigen synthetic peptides
According to the design of the antigen synthetic peptide, a phosphorylation group is added on a Ser131 site, and HPLC (high performance liquid chromatography) separation and purification are carried out, so that the purity of the obtained antigen synthetic peptide is more than 95%. Correspondingly, a synthetic peptide with a Ser131 site being non-phosphorylated is synthesized as a reference substance, and is coupled with hemocyanin (KLH for short) to form a polypeptide coupled protein which is used as a filler for affinity separation chromatography, and the purity of the polypeptide coupled protein after HPLC separation and purification is more than 90%.
EXAMPLE 3 preparation of antisera and purification of polyclonal antibodies
Animals were immunized (approximately 91 days in duration):
antigen treatment: one sterile syringe is used for sucking the antigen solution, the other syringe is used for sucking the equivalent amount of Freund's complete adjuvant (CFA), the two syringes are connected through a plastic tube, and the two syringes are repeatedly sucked back and forth until a completely emulsified emulsion is formed and is not dispersed when being dropped into water.
First immunization: the antigen emulsion is injected subcutaneously (s.c) at two sides of a part with thin and loose neck and back skin, muscle (i.m) at two sides of gluteus and thigh, intradermal (i.d) at two sides of waist, and multi-site multi-point injection such as rabbit paw pad, and the first immunization dose of the antigen is 1.4 mg.
First booster immunization: 14 days after the first immunization, the first booster immunization was performed, and antigen emulsion was prepared using Freund's incomplete adjuvant (IFA) instead of CFA as an immune adjuvant and injected in the first immunization manner, and the antigen dose of each booster immunization was 0.7 mg.
Second to fourth boosts: after 21 days of the last boosting immunization, IFA is used as an immunologic adjuvant to prepare antigen emulsion and is injected according to a first boosting immunization mode, and the last boosting immunization is carried out before a large amount of blood sampling.
After 14 days of the last boosting immunization, large blood sampling is carried out on the abdominal aorta of the white rabbit with big ear, and a large amount of antiserum is collected. The beaker with the collected blood was closed and allowed to stand overnight at room temperature to shrink the clot. The next day, aseptically packaging the separated serum into 50mL centrifuge tubes, centrifuging at 4000g for 10min, collecting supernatant, packaging 1mL each tube, labeling as immune antiserum, and storing at-20 deg.C.
Titer determination of antiserum after immunization: dropping the modified peptide and the control peptide as antigens on a cellulose acetate membrane at a ratio of 100 ng/drop and air-drying, sealing skim milk powder for 1 hour at room temperature, diluting antiserum (1: 1000, 1:5000, 1:10000, 1:50000, 1:100000, 1: 200000) by concentration gradient, detecting the binding titer of the antiserum and the antigens by Dot blot experiment (Dot blot), and detecting the result to show that the antiserum and the modified peptide have better affinity (the binding titer of the antiserum detected by the Dot blot is more than 1:50000) as shown in figure 2.
Purification of antiserum to polyclonal antibodies: after the antiserum is purified by using an affinity chromatography column, the polyclonal antibody, namely the polyclonal antibody phosphorylated by the serine at the 131 th position of BNIP3, is obtained.
Example 4 identification and use of polyclonal antibodies phosphorylated on serine 131 of BNIP3
1. Dot blob method is applied for identification
The dot blot experiment was performed using synthetic non-phosphorylated BNIP3 polypeptide (control peptide) and BNIP3 polypeptide having phosphorylation at serine 131 (modified peptide), which were dropped as an antigen at 100 ng/drop on a cellulose acetate membrane and air-dried, and then reacted with polyclonal antibody diluted with a concentration gradient. The results show that the polyclonal antibody phosphorylated on serine at position 131 of BNIP3 recognizes the polypeptide sequence phosphorylated on serine at position 131 of BNIP3, but does not recognize the non-phosphorylated BNIP3 polypeptide sequence, and the antibody titer >1:100000 (see fig. 3).
2. Identification by immunoblotting (Western Blot)
The wild-type pCDH-Flag-BNIP3-WT and mutant pCDH-Flag-BNIP3-S131/T137A expression vectors, a packaging vector psPAX2 and pMD2.G are co-transfected into HEK293T cells, cell culture supernatants are extracted to prepare viruses, the viruses are infected with PC12 cells and then screened by puromycin to finally construct two stable transfer cell strains of Flag-BNIP3-WT and Flag-BNIP3-S131/T137A, the two stable transfer cell strains and a non-infected Control (Control) PC12 cell precipitate are collected, and the phosphorylation level of IP3 is detected by an immunoblotting method after conventional lysis, so that the antibody can specifically recognize the BNIP3 phosphorylated at the 131 th serine (lane 2) and not recognize the non-phosphorylated S131/T137 (lane 3) 137A (lane 4).
3. Detection of exogenous over-expressed phosphorylated BNIP3 by immunoblotting (Western blot) method
Hela cells were cultured in DMEM medium containing 10% fetal bovine serum and grown in a 5% CO2 incubator. HeLa cells were transfected with the expression vector pCDNA3.1-Flag-BNIP3-WT of wild-type BNIP3 (FIG. 5 lane 2), the expression vector pCDNA3.1-Flag-BNIP3-S131A (FIG. 5 lane 3) of mutant BNIP3 and the empty vector pCDNA3.1 (FIG. 5 lane 1) by the conventional lipofection method, respectively, and the cells were collected after 48 hours of transfection and used for detection by the immunoblotting method. The results show that the antibody can specifically recognize exogenously overexpressed serine phosphorylated BNIP3 at position 131 (see fig. 5).
4. Detection of intracellular expressed phosphorylated BNIP3 by immunoblotting (Western blot) method
PC12 cells were placed in 20% O cells, respectively2Normoxic environment (lane 1 of FIG. 6) and 10% O2(FIG. 6 lane 2) and 0.3% O2(FIG. 6, lane 3) after culturing in two hypoxic environments for 24 hours, the phosphorylation level of BNIP3 in the cells was detected by immunoblotting after collecting the cells for lysis, and the result showed that the antibody specifically recognizes intracellularly expressed BNIP3 phosphorylated at serine 131 (see FIG. 6).
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (6)
1. An antigen synthetic peptide of a BNIP3 protein phosphorylation antibody, wherein the amino acid sequence of the antigen synthetic peptide is SSHCD (S-p) PPRSQ, and the phosphorylation site is serine at the 131 th position of BNIP3 protein.
2. A preparation method of a BNIP3 protein phosphorylation antibody is characterized by comprising the following steps:
1) coupling the antigen synthetic peptide of claim 1 with a carrier protein to obtain an antigen;
2) immunizing an animal by using the antigen obtained in the step 1), and collecting antiserum;
3) and (3) carrying out affinity column separation and purification on the antiserum obtained in the step 2) by using the synthesized modified peptide to obtain a Ser131 site phosphorylation antibody of the BNIP3 protein.
3. Use of the BNIP3 protein phosphorylating antibody of claim 2 for dot blot detection.
4. Use of the BNIP3 protein phosphorylating antibody of claim 2 for immunoblot detection.
5. Use of the BNIP3 protein phosphorylating antibody of claim 2 for detecting exogenous overexpression of phosphorylated BNIP3 in a cell.
6. Use of the BNIP3 protein phosphorylating antibody of claim 2 for detecting endogenous expression of phosphorylated BNIP3 in a cell.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010140528.4A CN111363031B (en) | 2020-03-03 | 2020-03-03 | pSer131 polyclonal antibody of BNIP3, preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010140528.4A CN111363031B (en) | 2020-03-03 | 2020-03-03 | pSer131 polyclonal antibody of BNIP3, preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111363031A true CN111363031A (en) | 2020-07-03 |
| CN111363031B CN111363031B (en) | 2022-03-11 |
Family
ID=71204279
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010140528.4A Active CN111363031B (en) | 2020-03-03 | 2020-03-03 | pSer131 polyclonal antibody of BNIP3, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111363031B (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 (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040152650A1 (en) * | 2002-07-22 | 2004-08-05 | Webster Keith A. | Preventing ischemia-induced cell damage |
| CN102492658A (en) * | 2011-12-15 | 2012-06-13 | 厦门大学 | PSer86 monoclonal antibody of Tip60, as well as preparation method and application of pSer86 monoclonal antibody |
| CN105585636A (en) * | 2015-11-24 | 2016-05-18 | 南方医科大学 | Human NOTCH1 NICD protein Ser2162 locus phosphorylation antibody and preparation method and application thereof |
| CN107298697A (en) * | 2017-08-24 | 2017-10-27 | 张灏 | Human PD-L1 protein Y123Site phosphorylation antibody and preparation method and application thereof |
| CN108752454A (en) * | 2018-06-19 | 2018-11-06 | 中山大学孙逸仙纪念医院 | A kind of human CYR 61 Protein S er188 site phosphorylations antigen, antibody and its preparation method and application |
| CN108840920A (en) * | 2018-06-19 | 2018-11-20 | 中山大学孙逸仙纪念医院 | A kind of human CYR 61 Protein S er167 site phosphorylation antigen, antibody and its preparation method and application |
-
2020
- 2020-03-03 CN CN202010140528.4A patent/CN111363031B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040152650A1 (en) * | 2002-07-22 | 2004-08-05 | Webster Keith A. | Preventing ischemia-induced cell damage |
| CN102492658A (en) * | 2011-12-15 | 2012-06-13 | 厦门大学 | PSer86 monoclonal antibody of Tip60, as well as preparation method and application of pSer86 monoclonal antibody |
| CN105585636A (en) * | 2015-11-24 | 2016-05-18 | 南方医科大学 | Human NOTCH1 NICD protein Ser2162 locus phosphorylation antibody and preparation method and application thereof |
| CN107298697A (en) * | 2017-08-24 | 2017-10-27 | 张灏 | Human PD-L1 protein Y123Site phosphorylation antibody and preparation method and application thereof |
| CN108752454A (en) * | 2018-06-19 | 2018-11-06 | 中山大学孙逸仙纪念医院 | A kind of human CYR 61 Protein S er188 site phosphorylations antigen, antibody and its preparation method and application |
| CN108840920A (en) * | 2018-06-19 | 2018-11-20 | 中山大学孙逸仙纪念医院 | A kind of human CYR 61 Protein S er167 site phosphorylation antigen, antibody and its preparation method and application |
Non-Patent Citations (3)
| Title |
|---|
| KATHERINE E LIU等: "Phosphorylation of the BNIP3 C-Terminus Inhibits Mitochondrial Damage and Cell Death without Blocking Autophagy", 《PLOS ONE》 * |
| UNKNOWN: "BNIP3 (Phospho-Ser95) antibody (HRP) orb503568", 《BIORBYT PRODUCT DATASHEET》 * |
| UNKNOWN: "RecName: Full=BCL2/adenovirus E1B 19 kDa protein-interacting protein 3", 《UNIPROTKB/SWISS-PROT: Q12983.3》 * |
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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111363031B (en) | 2022-03-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| de Silva et al. | Quality control in the endoplasmic reticulum: folding and misfolding of vesicular stomatitis virus G protein in cells and in vitro. | |
| US9506933B2 (en) | Method of preparing antigen for acquiring anti-hydrophobic peptide antibody | |
| US8569450B2 (en) | CD3 epsilon immunogens and antibodies | |
| NO854887L (en) | ANTIBODIES AGAINST HUMANT INTERLEUKIN-2 PROCEDURES OF SYNTHETIC POLYPEPTIDES. | |
| US5858723A (en) | Polypeptides and antibodies for diagnosing and treating seminoma | |
| CN111363031B (en) | pSer131 polyclonal antibody of BNIP3, preparation method and application thereof | |
| CN114058595B (en) | Hybridoma cell strain secreting anti-LAG 3 monoclonal antibody and application thereof | |
| EP0313156B1 (en) | Snrnp-a antigen and fragments thereof | |
| CN107298697B (en) | Human PD-L1 protein Y123Site phosphorylation antibody and preparation method and application thereof | |
| CN101735317A (en) | Cathepsin D antigen polypeptide and application thereof as well as detecting kit containing polypeptide | |
| CN107033225B (en) | Peste des petits ruminants virus HN protein epitope peptide and determination, preparation method and application thereof | |
| US20240018222A1 (en) | Monoclonal antibody inhibiting smim15 and application thereof | |
| CN101434653A (en) | Preparation and detection kit for human ARMET protein monoclonal antibody | |
| EP1094080A2 (en) | Anti-dnase-gamma antibody, and production and use thereof | |
| CN109651497B (en) | Grass carp FoxO1 polypeptide antigen and antibody, and preparation method and application thereof | |
| CN115290895A (en) | Application of methylation modification of 162 th lysine of human PD-L1 protein in prediction of immunotherapy sensitivity of malignant tumor | |
| CN106995801B (en) | Anti-tree shrew CD4 molecular monoclonal antibody, hybridoma cell strain secreting antibody and application | |
| US10308716B2 (en) | Method for producing polyclonal antibodies using an antigenic composition comprising protein-containing membrane vesicles | |
| CN117106058B (en) | Preparation method and application of pig neuromedin B receptor polypeptide and polyclonal antibody thereof | |
| CN106589123A (en) | Method for preparing anti-hyperglycosylation human chorionic gonadotropin antibody | |
| WO2023245800A1 (en) | Anti-cryptococcus capsular polysaccharide monoclonal antibody and use thereof | |
| CN116239684B (en) | Rabbit monoclonal antibody aiming at human calreticulin, and preparation method and application thereof | |
| CN108707186B (en) | Human sperm specific antigen epitope peptide, polymer and application thereof | |
| CN105315368B (en) | Anti-schistosoma japonicum Sj16 molecular monoclonal antibody and preparation method thereof | |
| CN115819521A (en) | Hybridoma cell strain secreting anti-cyprinid herpesvirus type 3 IL-10 monoclonal antibody, monoclonal antibody and application |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |