MilliporeSigma
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WB59

Millipore

Western-Re-Probe Reagent

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

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Quality Level

form

liquid

manufacturer/tradename

Calbiochem®

storage condition

OK to freeze

shipped in

wet ice

storage temp.

2-8°C

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.
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.

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 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

Legal Information

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

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 1

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


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