b'Restore Fluorescent Western Blot Stripping BufferThermo Scientific Restore Fluorescent Western Blot Stripping Buffer is a gentle and highly effective reagent for quickly removing primary antibodies and NIR dye-labeled secondary antibodies from western blots, allowing the same blot to be reprobed with different primary antibodies to detect alternative targets (Figure 52). Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membranes only.Features: Faststrip blots in only 15 minutes at room temperature Saves timeno need to run new gels and prepare aRestore Fluorescent LI-CORstripping bu\x0fer stripping bu\x0fernew blot and dyes and dyesMW 1 2 3 4 MW 1 2 3 4 Conserve samplesreprobe the same PVDF Initial blot membrane for multiple targets AKT 65 kDa (800) probed for Tubulin 55 kDa (700) AKT and tubulin Economicalless expensive than other commercially(channel 700 + 800)available stripping buffersAKT 65 kDa (800) Efficienteffectively strips blots the first time Tubulin 55 kDa (700) Stripped blot(channel 700 + 800)GAPDH 36 kDa (800) Stripped blotS6RP 30 kDa (700) reprobed for S6RP and GAPDH(channel 700 + 800)Figure 52. Effective stripping and reprobing of NIR fluorescent western blots on PVDF membranes. A549 whole cell lysates (20, 10, 5, and 2.5 g of protein in lanes 1, 2, 3, and 4, respectively) were transferred to PVDF membranes. Top panels: The blots were probed with AKT and tubulin antibodies and detected using Invitrogen DyLight 680 Goat AntiMouse IgG and DyLight 800 Goat AntiRabbit IgG, or IRDye 680 Goat AntiMouse IgG and IRDye 800CW Goat AntiRabbit IgG (LI-COR). An imaging system capable of fluorescence-based data capture (channel 700 and 800) was used for imaging. Middle panels: Blots were stripped with Restore Fluorescent Western Blot Stripping Buffer or NewBlot PVDF 5X Stripping Buffer (LI-COR) 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 antiS6 ribosomal protein (S6RP) and mouse anti-GAPDH antibodies. Targets were detected using DyLight 800 Goat AntiMouse IgG and DyLight 680 Goat AntiRabbit IgG, or IRDye 800CW Goat AntiMouse IgG and IRDye 680 Goat AntiRabbit IgG and imaged as described above.Learn more at thermofisher.com/westernbuffers74'