Western-Re-Probe Reagent

Description

• General description: Designed to dissociate and remove antibodies from membrane-bound proteins without destroying antigenic binding capacity. Recommended for stripping chemiluminescent or radioisotopic signals from blots. Not recommended for stripping color-producing Western blots. Supplied as a 5X solution sufficient for stripping 25-30 standard blots.
• Application: Immunoblotting (see comments)
• Components: 5X Re-Probe solution (125 ml)
• Warning: Toxicity: Standard Handling (A)
• Physical form: Supplied as a 5X solution.
• Other Notes: Sufficient volume to strip 25-30 standard blots.ADVANTAGES-Immunoblotting analyzed repeatedly with multiple probes-Saving on hard-to-get reagents and samples-Correction and re-analysis of unsatisfactory Western Blots-Saving time spent in gel re-running and transfer processesPROTOCOL1- Keep the membrane wet in an appropriate buffer after each probing procedure. Prepare 20-25 ml solution of 1X Western-Re-Probe in water.2- Incubate the membrane in 1X Western-Re-Probe buffer for 30 min at room temperature (RT) with shaking.3- Wash the membrane three times in PBST or TBST buffer containing 0.1% Tween-20 detergent. Use a large volume of washing buffer in a tray for the washing step. Each wash cycle involves agitating the membrane 5 min in the washing buffer.4- Block the membrane and perform immunodetection.Western-Re-Probe Reagent can be used for stripping and re-probing membranes 2-3 times. Some loss of signal may be noticed after second cycle of stripping and re-probing.PROTOCOL SUMMARY1. Incubate membrane in 1X Western-Re-Probe (30 min at RT) with shaking2. Wash the membrane 3 times in PBST or TBST buffer3. Block membrane and perform immunodetection
• Legal Information: CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany