Custom Premix Selecting "Custom Premix" option means that all of the beads you have chosen will be premixed in manufacturing before the kit is sent to you.
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.
A special formulation designed to dissociate and remove antibodies from the membrane bound protein without destroying the antigenic binding affinity.
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
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
17-July-2009 RFH
Application
Immunoblotting (see comments)
Description
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.
Form
Liquid
Formulation
Supplied as a 5X solution.
Comments
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
