Western Blot Stripping BufferBreaks antibody-antigen interactions to allow membranes to be reprobed using different antibodies

Western Blot Stripping Buffer

Western Blot Stripping Buffer is rated 4.4 out of 5 by 5.
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Application: Breaks antibody-antigen interactions to allow membranes to be reprobed using different antibodies
Supplemental Information: This is classified as a Dangerous Good for transport and may be subject to additional shipping charges.
* Refer to Certificate of Analysis for lot specific data (including water content).
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Western Blot Stripping Buffer is a gentle method for breaking antibody-antigen interactions to allow nitrocellulose and PVDF membranes to be reprobed several times using different antibodies, saving time and conserving samples. Ideal for use with chemiluminescent substrates. Used without dilution. One Western Blot Stripping Buffer package contains 500 ml Buffer A and 5 ml Buffer B, which combine to form sufficient reagent for approximately 25 membrane treatments.

Datasheet


References

1. Kabir-Salmani, M., et al. 2005. J. Clin. Endocrinol. Metab. 90: 4694-4702. PMID: 15886239

Usage :
•Wash already probed membrane with TBST (stock solutions available: sc-24953, sc-281695) for 5 min.

•Determine amount of stripping buffer needed (20 mL is usually sufficient for one membrane).

•Combine 20 mL of Buffer A and 200µL of Buffer B to make a 1:100 Buffer B: Buffer A dilution. For best results, let combined buffer come to room temperature prior to use.

•Incubate the membrane on an orbital shaker with sufficient stripping buffer to cover the membrane. Depending on the size and quantity, more than 20 mL may be needed.

•Incubate/shake for 30 min at RT. Results can be had in as little as 15 min, with up to 2 hrs for strongly bound antibodies.

•Wash 3x with TBST for 5 min.

•Reblock with appropriate blocking reagent, and perform western blot onto membrane according to Santa Cruz Biotechnology research applications or standard protocols.
Formulation :
Buffer A: Proprietary Component 1 40%, Water 60%
Buffer B: Proprietary component 2 10-20%, Water 80-90%
Physical State :
Liquid
Storage :
Store at 4° C
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.
Transport :
UN 3265, Class 8, Packing group III

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Certificate of Analysis

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Western Blot Stripping Buffer  Product Citations

See how others have used Western Blot Stripping Buffer. Click on the entry to view the PubMed entry .

Citations 1 to 2 of 2 total

PMID: # 26238153  Lian, F. et al. 2015. Mol Med Rep. 12: 5821-7.

PMID: # 23411318  Tsai, K.H. et al. 2013. Food chemistry. 138: 1856-67.

Citations 1 to 2 of 2 total

During the incubation with the stripping buffer, my membrane becomes transparent. Is this normal?

Asked by: Mathf
Thank you for your question. The change in appearance may be specific to what type of membrane you are using. We have observed some types of membranes become transparent after activation in methanol, so this is possible. Please contact us for Technical Support if you would like to provide more information about what type of membrane you are using,
Answered by: Technical Support
Date published: 2019-06-04

Can you use this buffer for nitrocellulose or pvdf?

Asked by: woojonglee0917
Thank you very much for the question. Yes, our Western Blot Stripping Buffer is a gentle method for breaking antibody-antigen interactions to allow both, nitrocellulose and PVDF membranes, to be re-probed several times using different antibodies, saving time and conserving samples. THis products is ideal for use with chemiluminescent substrates. 
Answered by: Technical Support Europe
Date published: 2019-01-28

What does sc-281698 contain?

Asked by: chemicalsmg
Thank you for your question. Our product Western Blot Stripping Buffer, sc-281698, is a two vial kit. The following chemicals are present: Guanidine Hydrochloride CAS 50-01-1; Tris base CAS 77-86-1; TCEP; Hydrochloride CAS 51805-45-9; Water CAS 7732-18-5. The formulation is currently proprietary.
Answered by: Chemical Support 3
Date published: 2017-03-01

Can you reuse the stripping buffer for western blots? If so, how many times?

Asked by: chemicalsmg
Thank you for your question. For our product Western Blot Stripping Buffer, sc-281698, we recommend preparing fresh A+B Buffer when stripping a new blot.
Answered by: Chemical Support 3
Date published: 2017-03-01

Are either buffers a phosphate based buffer?

Asked by: chemicalsmg
Thank you for your question. For our product Western Blot Stripping Buffer, sc-281698, Buffer B contains Tris (2-carboxyethyl) phosphine (TCEP).
Answered by: Chemical Support 3
Date published: 2017-03-01

Does the product contain any reducing agents?

Asked by: chemicalsmg
Thank you for your question. Western Blot Stripping Buffer, sc-281698, contains Tris (2-carboxyethyl) phosphine (TCEP).
Answered by: Chemical Support 3
Date published: 2017-03-01
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Rated 5 out of 5 by from Excellent for stripping the western blots I bought last month and I wasn’t sure if it’s going to work but it give me excellent results
Date published: 2018-05-15
Rated 5 out of 5 by from Good product We purchased it last months it worked really nicely, i recommend it strongly.
Date published: 2017-06-08
Rated 5 out of 5 by from works well we used this product first time and surprised with the outcome, works well
Date published: 2017-06-06
Rated 2 out of 5 by from Mismatching size! Our lab use this stripping buffer for a long time, but there is one thing perplexes us much which is the buffer B (the small one) is too little to pair with buffer A (the big one). We always run out of buffer B before we actually finish the buffer A which means we always need to buy a pair of brand new stripping buffer just for buffer B! It's so annoying though. One suggestion, the company could offer buffer A and B separately for the customers or, why just add up the size of buffer B. The current design of the size is not reasonable, truly.
Date published: 2017-03-17
Rated 5 out of 5 by from Various publications cite the use of western Various publications cite the use of western blot stripping buffers in the Western Blotting applications in order to re-probe their membrane for another antibody. -SCBT Publication Review
Date published: 2015-07-15
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