RenovaStrip™ Stripping Buffer

CatNo.:IC-8013

Description

RenovaStrip™ Stripping Buffer provides a robust but gentle method for removing primary and secondary antibodies from western blots that were detected with HRP substrates. RenovaStrip™ Stripping Buffer allows forreprobing, saving time and costs when samples are in limited quantities, when the same sample requires analysis by different antibodies,and when optimization is required. RenovaStrip™ Stripping Buffer can be used with both nitrocellulose(NC) and PVDF membranes.

Size

100ml.500ml

Important Guidelines

1.RenovaStrip™ Stripping Buffer is supplied in ready-to-use format. It is not recommended to dilute the buffer.

2.Membranes may be stored in TBST at 4¡æ until the stripping procedure can be performed.

3.RenovaStrip™ Stripping Buffer will not dissociate interactions between a biotinylated target protein and avidin-conjugated probes.

4.Stripping fluorescent western blots is not recommended with RenovaStrip™ Stripping Buffer as results can be inconsistent. It is recommended to use RenovaStrip™Fluorescent Western Blot Stripping Buffer for fluorescent applications.

5.Blocking the membrane is recommended after stripping.

Usage Protocol

1. Bring RenovaStrip™ Stripping Buffer to room temperature.

2. Wash blot once for 5 minutes in wash buffer (TBST) to remove the HRP substrate.

3. Add sufficient RenovaStrip™ Stripping Buffer to cover the blot and incubate for 15 minutes at 37¡æ with gentle shaking.

Note: Optimization of both incubation time and temperature is essential for best results. For some antibodies, room temperature incubation is sufficient. However, high-affinity antibodies or saturated blots (excess secondary antibody) may require incubation for an additional 15 to 30 minutes at 37¡ãC.

4. Remove the blot from the RenovaStrip™ Stripping Buffer and wash 3 times for 5 minutes each in TBST with gentle shaking.

5. Test the effectivenss of stripping .incubate the membrane with new HRP substrate. If no signal is detected using a 5-minute exposure, the HRP conjugate has been successfully removed from the antigen or primary antibody. To test for complete removal of the primary antibody, incubate the membrane with the HRP-labeled secondary antibody,followed by 5-minute washes in wash buffer. Incubate the membrane with new HRP substrate. If no signal is detected with a 5 minutes exposure, the primary antibody has been successfully removed from the antigen.

6. If signal is detected with either test in step 5, incubate blot in RenovaStrip™Western Blot Stripping Buffer for an additional 5 to 15 minutes and repeat step 3 and step 4.

Note: Some antigen/antibody systems require increased temperature and/or longer incubation times to strip them fully. Optimize stripping time and temperature to ensure complete removal of antibodies while preventing damage to the antigen.

7. After determining that the membrane is properly stripped, the second immunoprobing experiment may be performed.

8. Block the membrane in blocking buffer before continuing with the second immunoprobing experiment.

Storage

Store at 2-8 ¡æ. 

Price