Portal de Periódicos da CAPES

Reutilization of Immunoblots after Chemiluminescent Detection

2001; Elsevier BV; Volume: 296; Issue: 2 Linguagem: Inglês

10.1006/abio.2001.5313

1096-0309

Scott H. Kaufmann,

Advanced Biosensing Techniques and Applications

Inactivation of horseradish peroxidase (HRP) treatment is a conventional preference to stripping for sequential detections of different proteins of chemiluminescent western blotting (WB). However, little evidence exists on whether other chemical substances treatment can affects the biological activity of HRP during stripping and re-probing of WB blots. Here, we successfully develop 20% crotonic acid (CA) as an alternative to stripping to inhibit HRP used for sequential chemiluminescent WB on polyvinylidene difluoride (PVDF) and Nitrocellulose (NC) membrane. Moreover, NC blots incubation in CA (40 °C, 30min) allow us to perform three round HRP inhibition in sequential detections without losing transferred proteins and damaging membrane. Hence, the method will help us save time and valuable samples without the need to rerun gels.