Re-use Old or New Western Blots using ADI’s Western Blot Recycling Kit
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Strip antibodies at RT; No heating required |
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Just soak blots in stripping solution for 10-15 min |
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No mercaptoethanol; No pungent smell |
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Blots are reusable within 15-60 min |
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Blots can be probed with the same or different antibodies |
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Western Blot Recycling Kit Cat #90100 Sufficient for 30-50 Mini-Blots |
Western blotting is a commonly used technique to study protein structure and functions. Typically, protein samples are electrophoresed on SDS-PAGE and transferred to solid support (nitrocellulose or nylon-based membranes) for probing with antibodies. Unlike advances made in similar techniques in probing nucleic acids (northern and southern blotting) that allow reuse of blots, it has been difficult to reuse blots for Western blotting. Recycling of protein blots offers many advantages:
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- When protein samples are available in limited quantities, difficult to obtain or expensive
- When samples are to be analyzed with different antibodies under identical conditions, e.g., subtype or isoform specific antibodies
- When a blot gives unexpected results and needs confirmation with the same or a different antibody
- When a blot is mistakenly incubated with wrong antibody
- It is simply more economical and less time consuming to reuse the same blot.
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List of publications: |
Humphreys 2002, Endocrinology 143, 3641| Used Nitrocellulose| Stripped 4 times and reprobed with other antibodies
Dwivedi 2002, JBC 277, 29643, Used Nitrocellulose, Stripped phosphoantibodies and reprobed with other antibodies
McClelland 2002, J Appl Physiol, 2002| 92: 1573, Used PVDF, stripped and reprobed with various MCTs antibodies
Jackson 2001 , J Immunol 166, 952-958| Used Nitrocellulose| primary antibody stripped and reprobed with beta-actin |
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