Restore™ Western Blot Stripping Buffer

Thermo Scientific Restore Western Blot Stripping Buffer safely and effectively removes primary and secondary antibodies from nitrocellulose and PVDF membranes to allow chemiluminescent Western blots to be reprobed.

Features of Restore Western Blot Stripping Buffer:

• Saves time—no need to re-run gels and blots
• Saves costly sample—re-probe the membrane using the same target sample
• Effective—formulation is more efficient at stripping antibodies than homemade buffers
• Gentle—does not damage the target antigen during stripping allowing efficient reprobing
• Odor-free—no mercaptans means no acrid odors
• Economical—less expensive than other commercial stripping buffers

Product Details

Performing gel electrophoresis and duplicate immunoblot assays to test new primary antibodies or antibody concentrations is time-consuming and expensive. Restore Western Blot Stripping Buffer eliminates this waste when detecting immunoblots with chemiluminescent Western blotting substrates. Restore Stripping Buffer provides clean and efficient removal of primary and secondary antibodies from immunoblots without removing or damaging the immobilized antigen allowing blots to be stripped and reprobed with confidence.

Chemiluminescent Western blot detection with reagents such as Thermo Scientific SuperSignal Substrates for horseradish peroxidase is one of the most common and sensitive methods in use today. Because these substrates do not precipitate and bind to membrane surfaces, Western blots detected by chemiluminescence can be stripped with reagents that remove affinity-bound primary and secondary antibodies. To be effective, a stripping buffer must be strong enough to disassociate bound antibodies but gentle enough to leave the transferred target proteins intact on the nitrocellulose or PVDF membrane. Restore Western Blot Stripping Buffer has these characteristics.

By stripping and reprobing, there is no need to waste rare or costly samples by running multiple gels in order to probe for different targets. A single membrane from one gel can be stripped with Restore Western Blot Stripping Buffer to remove the primary antibodies. Stripping the blot takes only 15 to 30 minutes, depending on the affinity of the primary antibody. After stripping, block and reprobe with a new primary antibody. Alternatively, a blot can be stripped and reprobed with adjusted antibody concentrations to optimize conditions after obtaining initially poor results.

Applications

• Reuse a nitrocellulose or PVDF blot to detect a different target with a different primary antibody
• Reprobe a blot to correct or optimize antibody concentrations that were ineffective the first time

Protocol Summary

• Wash blot to remove chemiluminescent substrate.
• Incubate blot in Restore Western Blot Stripping Buffer for 5 to 15 minutes at 37°C (room temperature is sufficient for some antibodies).
• Remove blot and wash in Wash Buffer (TBS-T or PBS-T).
• Test for sufficient removal of antibodies.
• Perform next immunoblot experiment.