title: [Preparation and identification of rabbit anti-AKR1B10 polyclonal antibody].
abstract: Objective To prepare rabbit anti-aldo-keto reductase family 1 member B10 (AKR1B10) polyclonal antibody and identify its specificity. Methods AKR1B10 cDNA was amplified by reverse transcription PCR (RT-PCR) and inserted into prokaryotic expression vector pET-15b to form recombinant plasmid pET-15b-AKR1B10. The recombinant plasmid pET-15b-AKR1B10 was transformed into E.coli DH5α. Isopropylthio-β-D-galactoside (IPTG) was used to induce the expression of the recombinant protein His-tagged AKR1B10 in E.coli DH5α. The expression products from different clones of E.coli DH5α were identified by SDS-PAGE. The positive bacteria were picked out and amplified. His-Tag-AKR1B10 protein was purified from the expression product of the positive clones by His-tagged purification column. The purified recombinant protein His-Tag-AKR1B10 was used to immunize New Zealand white rabbits. Antisera were acquired after two months. Anti-AKR1B10 polyclonal antibodies were purified by antigen purification column with AKR1B10 recombinant protein. Lastly, the purified polyclonal antibodies were identified by SDS-PAGE, ELISA, Western blotting. Results The recombinant plasmid pET-15b-AKR1B10 was constructed successfully, and the recombinant protein His-Tag-AKR1B10 with high purity was acquired. The purified polyclonal antibodies were able to specifically recognize AKR1B10 protein. Conclusion The rabbit anti-AKR1B10 polyclonal antibodies is prepared successfully with high specificity.
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China; Center for Clinical Pathological Laboratory, Chenzhou 423000, China.
authors: Jian
authors: J
authors: Hu
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China; Center for Clinical Pathological Laboratory, Chenzhou 423000, China.
authors: Qingmei
authors: Q
authors: Wang
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China; Center for Clinical Pathological Laboratory, Chenzhou 423000, China.
authors: Li
authors: L
authors: Huang
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China; Center for Clinical Pathological Laboratory, Chenzhou 423000, China.
authors: Yuanqing
authors: Y
authors: Zeng
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China.
authors: Zheng
authors: Z
authors: Hu
authors: National & Local Joint Engineering Laboratory for High-through Molecular Diagnosis Technology, Affiliated First People's Hospital of Chenzhou City, University of South China; Translational Medicine Institute, University of South China; Center for Clinical Pathological Laboratory, Chenzhou 423000, China. *Corresponding author, E-mail: luodixian_2@163.com.
authors: Dixian
authors: D
authors: Luo
mesh_terms: D000449:Aldehyde Reductase / Q000276:immunology*; D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000378:metabolism*; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D007544:Isopropyl Thiogalactoside; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D007544:Isopropyl Thiogalactoside; D000449:Aldehyde Reductase