Western Blot Stripping Buffer (StripSure™)

BioFargo Inc. · Ready-to-use · 500 mL Western Blot Stripping Buffer (StripSure™) Mild, ready-to-use antibody stripping buffer for complete removal in 20 minutes at room temperature—without heat or harsh chemicals. StripSure™ efficiently removes primary and secondary antibodies from Western blot membranes while preserving membrane-bound target proteins for reliable re-probing. It is compatible with PVDF, nitrocellulose, and nylon membranes; simply immerse, incubate, rinse, and re-probe. Performance testing confirmed removal of detectable signal from previously probed antibodies (anti-ATG7), while re-probing generated strong, specific signal with β-actin and phospho-STAT3 (Tyr705). Its gentle conditions support sensitive phospho-specific detection workflows. 20 minRoom temperature protocol MildNo heat or reducing agents 3 membranesPVDF, nitrocellulose & nylon Key Features & Benefits Efficient removal of primary and secondary antibodies Compatible with phospho-specific antibody re-probing Helps preserve membrane-bound target proteins across stripping cycles Ready-to-use liquid formulation—do not dilute 20-minute protocol at room temperature (20–25°C) with gentle agitation Suitable for chemiluminescent and fluorescent detection systems Mild vs. Harsh Stripping: Which Do You Need? Stripping buffers fall into two categories, and choosing the wrong one is a common reason a re-probe fails. Harsh stripping uses SDS and β-mercaptoethanol at 50–70°C. It removes essentially any antibody, including high-affinity antibodies, but can also strip target protein from the membrane. Signal loss on a second probe can be measurable, particularly for low-abundance targets. Mild stripping uses low pH and mild detergent at room temperature. It removes antibody without displacing membrane-bound protein, supporting sequential probing that is quantitative rather than qualitative. StripSure is a mild stripping buffer. It requires no heat, β-mercaptoethanol, or reducing agents: 20 minutes at room temperature. Use it when the second blot must be comparable to the first, when re-probing phospho-epitopes, or when a low-abundance target cannot tolerate signal loss. If you are stripping a very high-affinity antibody and preserving the target is not important, a harsh protocol may still be faster. For other applications, mild stripping is the appropriate default. Western Blot Stripping Protocol (20 Minutes, Room Temperature) Step-by-Step Initial wash — Rinse the membrane once with deionized water to remove residual detection reagents. Stripping — Gently mix StripSure™ until homogeneous (do not dilute), then completely immerse the membrane. Incubation — Incubate for 20 minutes at room temperature (20–25°C) with gentle agitation. Rinsing — Rinse three times with deionized water, or three times with TBST for sensitive applications. Re-equilibration — Equilibrate the membrane in TBST for 5 minutes. Re-probing — Proceed with primary antibody incubation, wash, secondary antibody incubation, wash, and detection. Important: Keep the membrane fully submerged throughout stripping for consistent, even results. Stripping and Re-Probing Phospho-Specific Blots Phospho-specific targets are particularly vulnerable to signal loss under heated or reducing stripping conditions. StripSure’s room-temperature, non-reducing mild stripping protocol is intended to retain membrane-bound protein for subsequent phospho-specific re-probing. Troubleshooting: When Stripping Doesn’t Work Observation Most likely cause What to do Original signal is still visible High-affinity antibody or membrane not fully immersed Run another 20-minute cycle; ensure full immersion and continuous gentle agitation. Second-probe signal is much weaker Target protein was eluted, often after a prior harsh protocol Use a mild protocol; for low-abundance targets, consider re-running the blot. High background on re-probe Insufficient washing or residual stripping buffer Wash with TBST three times for 5 minutes each; re-block for at least 1 hour. Blotchy or uneven signal Membrane dried during incubation Keep it wet throughout; move directly to washing after stripping. Phospho signal is lost A heated or reducing protocol was used Use a room-temperature, non-reducing mild protocol for phospho-epitopes. Nothing works after two cycles The membrane reached its stripping limit Do not strip the same membrane more than 2–3 times; re-run the blot. StripSure™ vs. Restore™ PLUS StripSure™ Restore™ PLUS (46430) 500 mL price $120 $189.65 Incubation conditions 20 min, room temperature 5–15 min, room temperature; 37°C for high-affinity antibodies Heat required No 37°C for difficult-to-strip antibodies Membrane compatibility PVDF, nitrocellulose, nylon Nitrocellulose, PVDF Ready to use Yes; no dilution Yes Pricing and specifications as published by the manufacturer, August 2026. Specifications Product Name StripSure™ Western Blot Stripping Buffer Brand BioFargo Inc. Format Ready-to-use liquid (do not dilute) Compatible Membranes PVDF, nitrocellulose, nylon Detection Systems Chemiluminescent and fluorescent imaging Incubation 20 minutes at room temperature (20–25°C), gentle agitation Antibody Compatibility Primary and secondary antibodies, including phospho-specific antibodies Rinse Options Deionized water or TBST (recommended for sensitive applications) Size / Volume 500 mL bottle Storage Room temperature Shelf Life 1 year Validation Data StripSure™ was validated for antibody removal using anti-ATG7 and for re-probing using β-actin and phospho-STAT3 (Tyr705). Western blot validation data Stripping and re-probing validation data Frequently Asked Questions How long does Western blot stripping take? StripSure requires a 20-minute incubation at room temperature with gentle agitation. Can you strip and re-probe a phospho-specific Western blot? Yes. StripSure is a mild, room-temperature, non-reducing buffer designed for re-probing, including phospho-specific workflows. How many times can you strip the same membrane? Typically no more than two to three times. Re-run the blot after that. Is StripSure a mild or harsh stripping buffer? StripSure is a mild stripping buffer; it works at room temperature without heat, β-mercaptoethanol, or reducing agents. Does it work with nitrocellulose and PVDF? Yes. It is compatible with PVDF, nitrocellulose, and nylon membranes. Do I need to re-block after stripping? Yes. After stripping and washing, re-block the membrane for at least one hour before re-probing. What is the difference between mild and harsh stripping buffers? Harsh buffers use SDS and β-mercaptoethanol at elevated temperatures. Mild buffers use low pH and mild detergent at room temperature to help preserve membrane-bound protein. Download Product User Manual