ReBlot Western Blot Recycling Kit
Western blotting is a commonly used technique for studying protein function and localization. Typically, protein samples are electro-phoresed on SDS-PAGE and transferred to a membrane such as nitrocellulose or nylon, where they are probed with specific
antibodies. Unlike nucleic acid based technologies, which allow reuse of Southern and Northern blots, it has been difficult to reuse Western blots.
Stripping and re-probing of Western blots offers several advantages:
1) Conservation
of samples that are expensive or available only in limited quantities,
2) Analysis of a given blot using several different antibodies, e.g. subtype- or isoform-specific antibodies,
3) Re-analysis of anomalous results and confirmation
with the same or a different antibody,
4) Correcting errors in incubation with the wrong antibody,
5) Cost savings in reagents and time by reusing the same blot.
While antigen and antibody-based immunoaffinity matrices,
such as Sepharose™ conjugates, have been reused many times without compromising antigen-antibody reactivity, the need for pH extremes and chaotropic agents has precluded the application of these methods to Western blotting.
The MILLIPORE
Re-Blot Western Blot Recycling Kit contains specially formulated solutions that quickly and effectively remove antibodies from Western blots without significantly affecting the immobilized proteins.
Advantages of the Re-Blot Western Blot
Recycling Kit include:
·*No pungent-smelling b-mercaptoethanol is contained in the Antibody Stripping Solution.
·*Antibody stripping is done at room temperature. No heating of blots is required.
·*Blots can be stripped
of antibodies in approximately 10 minutes at room temperature.
·*Reblocking of blots may be avoided in most instances.
·*Blots may be reused in less than 15 minutes. |
| Materials Required but Not Delivered |
· Standard Blot or blot strips, on nitrocellulose or PVDF/nylon membrane.
· Alternative Blocking Solutions, such as 10% BSA, for antibodies that are not compatible with milk-based blotting agents.
· Plastic Wrap, such as Saran Wrap,
for storage of blots that will not be re-probed immediately.
· Distilled Water, for reagent dilution.
· Plastic Trays for incubation of blots or blot strips in stripping, washing and blocking solutions.
· Positive and Negative
Stripping Controls. It is recommended that one new strip (not subjected to stripping solution) be included for comparison purposes. |
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