Preparation and application of rabbit anti-FAM50B polyclonal antibody
Objective To prepare rabbit anti-FAM50B polyclonal antibody and investigate the application of FAM50B's function.Methods The expression and prognosis of FAM50B in seven cancers were analyzed using the GEPIA online platform.The pGEX-6P-1-FAM50B prokaryotic expression plasmid was constructed utilizing DNA recombination technology and transformed into E.coli BL21.Expression was induced with different IPTG concentrations,and the FAM50B fusion protein was purified using a GST purification column.The purified target protein was used for immunization experiments in New Zealand rabbits.The specificity of the homemade FAM50B polyclonal antibody was evaluated using Western Blot,while its localization and expression in mouse tissues were assessed through immunofluorescence staining.Results FAM50B is abnormally expressed in seven types of cancer,and its expression is associated with poor prognosis in LGG.The pGEX-6P-1-FAM50B plasmid was successfully constructed,and significant expression of the FAM50B fusion protein,approximately 62 kDa in size,was observed at 16℃ after 16 hours of induction with 0.3 mM IPTG.The homemade antibody demonstrated good specificity,with high and specific expression of FAM50B in the testes,localized in the cell nucleus,as shown by Western Blot and immunofluorescence staining.Conclusion The prepared FAM50B antibody serum exhibits high specificity and potency,suitable for Western Blot and immunofluorescence staining,thus establishing a solid basis for further in-depth studies on FAM50B's function.
FAM50BRabbit antibodyProkaryotic expressionProtein purificationBioinformatics analysisAntibody preparation and application
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