Western-Re-Probe Reagent

Immunoblotting (see comments)

Toxicity: Standard Handling (A)

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.

This Western-Re-Probe Reagent is validated for use in Immunoblotting.

Western-Re-Probe Reagent is specifically formulated to dissociate and remove antibodies from the membrane bound protein without destroying the antigenic binding affinity. The removal of antibodies also removes corresponding chemiluminescence or radioactive signals from the blots. Stripped blots can be probed with new probes.

5X Re-Probe solution (125 ml)

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 processes



PROTOCOL

1- 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 SUMMARY

1. Incubate membrane in 1X Western-Re-Probe (30 min at RT) with shaking

2. Wash the membrane 3 times in PBST or TBST buffer

3. Block membrane and perform immunodetection

Supplied as a 5X solution.

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany