Restore Fluorescent Western Blot Stripping Buffer

The Thermo Scientific Restore Fluorescent Western Blot Stripping Buffer is a gentle and highly effective reagent for quickly removing primary and near-infrared (IR) dye-labeled secondary antibodies from Western blots.

Restore Fluorescent Western Blot Stripping Buffer enables the reuse of PVDF membranes, simplifying the Western blot optimization process and allowing the same blot to be reprobed with different primary antibodies to detect alternative targets. Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membrane only (Part No. 22860).

Highlights:

• Fast – strip blots in only 15 minutes at room temperature
• Saves time – no need to run new gels and prepare a new blot
• Conserve samples – reprobe the same PVDF membrane for multiple targets
• Economical – less expensive than other commercially available stripping buffers
• Efficient – effectively strips blots the first time

Product Details:

Fluorescence Western blotting is a powerful method for detecting multiple targets at once. Restore Fluorescent Western Blot Stripping Buffer allows re-probing of PVDF membranes, saving time and cost. This is ideal when samples are limited and optimization or analysis with different primary antibodies is required.

Traditional stripping methods may adversely alter or remove the sample proteins from the PVDF membrane during the stripping process or may be effective for removing only low-affinity antibodies. In contrast, Restore Fluorescent Western Blot Stripping Buffer Stripping efficiency exceeds 90% while reprobing efficiency matches or exceeds other supplier’s formulations. Also, Restore Fluorescent Western Blot Stripping Buffer is conveniently stored at room temperature and easy to use. Simply dilute buffer 1:5 in water and incubate your membrane for 5 to 20 minutes at room temperature.

Effective stripping and reprobing of near-IR fluorescent Western blots on PVDF. A549 whole cell lysates (20, 10, 5, and 2.5µg of protein in lanes 1, 2, 3, and 4, respectively) were transferred to PVDF membranes. Top panels: The blots were probed with anti-Akt and anti-tubulin antibodies and detected using DyLight 680 Goat Anti-Mouse IgG (Part No. 35519) and DyLight 800 Goat Anti- Rabbit IgG (Part No. 35571), or IRDye* 680 Goat Anti-Mouse and IRDye 800CW Goat Anti-Rabbit (LI-COR Biosciences). The LI-COR Odyssey* Infrared Imaging System (channel 700 and 800) was used for imaging. Middle panels: Blots were stripped with Restore Fluorescent Stripping Buffer or NewBlot* PVDF Stripping Buffer for 15 minutes at room temperature. Blots were rinsed with TBS for 5 minutes and re-imaged using channel 700 and 800. Bottom panels: Stripped membranes were reprobed with rabbit anti-S6 ribosomal protein and mouse anti-GAPDH antibodies. Targets were detected using DyLight 800 Goat Anti- Mouse IgG (Part No. 35521) and DyLight 680 Goat Anti-Rabbit IgG (Part No. 35568), or IRDye 800CW Goat Anti-Mouse and IRDye 680 Goat Anti-Rabbit and imaged as described above.

Related Products: