Author: Kim, Nam-Gun; Kim, Myeong-Ae; Park, Young-Il; Jung, Tae-Sung; Son, Seong-Wan; So, ByungJae; Kang, Hwan-Goo
Title: Magnetic nanoparticle based purification and enzyme-linked immunosorbent assay using monoclonal antibody against enrofloxacin Document date: 2015_12_17
ID: 5i84nomn_8
Snippet: The immunogen ENR-KLH and coating antigen ENR-BSA were prepared using the NHS ester method as previously described, with slight modifications [15] . In this procedure, a total of 4 mg of ENR (0.011 mmol) was dissolved in DMF (35 µL) and added to 0.25 MNHS solution (40 ïL) and EDC (40 mg, 0.258 mmol) with continuous stirring. The mixture was then incubated for 24 h at 4 o C without light, after which 2.5 mL of coupling buffer with 10% (w/w) KLH.....
Document: The immunogen ENR-KLH and coating antigen ENR-BSA were prepared using the NHS ester method as previously described, with slight modifications [15] . In this procedure, a total of 4 mg of ENR (0.011 mmol) was dissolved in DMF (35 µL) and added to 0.25 MNHS solution (40 ïL) and EDC (40 mg, 0.258 mmol) with continuous stirring. The mixture was then incubated for 24 h at 4 o C without light, after which 2.5 mL of coupling buffer with 10% (w/w) KLH or BSA were slowly added to the mixture solution while stirring, followed by 6 h of incubation at 4 o C without light. Finally, the reaction mixture was dialyzed (molecular weight cut-off : 10,000 Da) against PBS (0.1 mol/L, pH 7.4) at 4 o C and stored at −20 o C until use as the immunogen. The UV absorbance method was used to determine if the conjugate was a success.
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