In Western Blot (WB) experiments, antibody stripping is mainly used in the following scenarios, depending on experimental needs and antibody properties:
1. Detecting multiple targets on the same membrane
When sample quantity is limited or consistent detection conditions are required, stripping allows membrane reuse. Examples include:
1) Sequential detection of different targets: Detect low-abundance proteins first, then strip and detect high-abundance ones to avoid incomplete stripping.
2) Multi-marker validation: For example, detecting proteins in the same signaling pathway (e.g., phosphorylated and total proteins). Incubate with the phospho-antibody first, then strip and probe for total protein to ensure data comparability.
2. Optimizing antibody incubation or verifying specificity
1) Exclude non-specific binding: If unexpected bands appear, stripping and reprobing with a different antibody can help confirm cross-reactivity.
2) Adjust antibody order: Detect weak or easily stripped antibodies first, followed by stronger ones.
3. Saving sample and time
1) Limited sample availability: For rare or precious samples (e.g., clinical or primary cells), reusing the membrane reduces sample usage.
2) Faster experimental turnaround: Stripping is quicker than redoing gel and transfer, ideal during optimization.
4. Special experimental needs
1) Loading control correction: Detect target and reference proteins sequentially on the same membrane to avoid batch variation.
2) Blocking adjustment: If BSA impairs stripping efficiency, switch to Servicebio's protein-free quick blocker or skim milk to improve results.
Notes:
Stripping limit: A membrane can typically be stripped 3–4 times; excessive stripping may result in protein loss.
Effectiveness check: Always verify antibody removal by ECL detection to avoid false positives.
In summary, antibody stripping is ideal for multi-target detection, result validation, and sample conservation. Choose stripping conditions based on your experiment and antibody properties.
We recommend Servicebio Antibody Stripping Buffer for safe and efficient removal of primary and secondary antibodies from PVDF or NC membranes:
G2078 (Regular): For weak signals or low-affinity antibodies.
G2079 (Enhanced): For strong signals or high-affinity antibodies.
