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| Application of quantitative dot blot analysis technology for protein expression level detection |
| LIU Lin1, MEN Tingting2, ZHANG Wenfeng3, ZHANG Jiandi3, MI Jia4, YANG Chunhua4* |
1 Department of Pharmacy, Binzhou Medical University, Yantai 264003 Shandong,P.R.China;
2 Department of Nursing, Binzhou Medical University;
3 Yantai Zestern Biotechnique Co. LTD;
4 Medicine and Pharmacy Research Center, Binzhou Medical University |
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Abstract Objective To detect the application of quantitative dot blot analysis (QDB analysis) detection technology for protein expression level analysis by determining the protein level of high throughput tissue samples and cell samples. Methods The relative expression of tubulin in the prostate tissues from 87 mice and the absolute expression of tubulin from 8 human cell lines were detected by QDB analysis. Results The results showed that the expression level of tubulin protein in prostate tissue from mice was different according to the age, and the absolute expression level of tubulin protein in 8 cell lines was between 90～140 fmol/μg. Conclusion QDB analysis can be applied to the relative and absolute quantification in a high throughput tissue samples and cell samples.
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Received: 05 December 2019
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[1] RENART J, REISER J,STARK G R. Transfer of proteins from gels to diazobenzyloxymethyl-paper and detection with antisera: a method for studying antibody specificity and antigen structure [J]. Proc Natl Acad Sci U S A, 1979, 76(7):3116-3120.
[2] BURNETTE W N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A [J]. Anal Biochem, 1981, 112(2):195-203.
[3] ENGVALL E,PERLMANN P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G [J]. Immunochemistry, 1971, 8(9):871-874.
[4] ENGVALL E, JONSSON K, PERLMANN P. Enzyme-linked immunosorbent assay. II. Quantitative assay of protein antigen, immunoglobulin G, by means of enzyme-labelled antigen and antibody-coated tubes [J]. Biochim Biophys Acta, 1971, 251(3):427-434.
[5] QI X, ZHANG Y, ZHANG Y, et al. High throughput, absolute determination of the content of a selected protein at tissue levels using quantitative Dot Blot analysis (QDB) [J]. J Vis Exp, 2018(138): e56885.
[6] TIAN G, TANG F, YANG C, et al. Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method [J]. Oncotarget, 2017, 8(35):58553-58562.
[7] GREENBERG N M, DEMAYO F, FINEGOLD M J, et al. Prostate cancer in a transgenic mouse [J]. Proc Natl Acad Sci U S A, 1995, 92(8):3439-3443.
[8] TOGTEMA M, HUSSACK G, DAYER G, et al. Single-Domain Antibodies Represent Novel Alternatives to Monoclonal Antibodies as Targeting Agents against the Human Papillomavirus 16 E6 Protein [J]. International Journal of Molecular Sciences, 2019, 20(9):pii: E2088.
[9] BYWATERS S M, BRENDLE S A, TOSSI K P, et al. Antibody Competition Reveals Surface Location of HPV L2 Minor Capsid Protein Residues 17-36 [J]. Viruses, 2017, 9(11): pii: E336. |
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