Thermo Scientific Restore PLUS Western Blot Stripping Buffer is an advanced formula for removing bound primary and secondary antibodies fromRead more
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Quantity
46428
30 mL
46430X4
4 x 500 mL
46430
500 mL
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Catalog number 46428
Price (USD)
56.50
30 mL
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Quantity:
30 mL
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Price (USD)
56.50
30 mL
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Thermo Scientific Restore PLUS Western Blot Stripping Buffer is an advanced formula for removing bound primary and secondary antibodies from membranes so they can be reprobed and detected with chemiluminescent substrates.
Features of Restore PLUS Western Blot Stripping Buffer:
• Ready and easy to use—no dilution necessary; no offensive odors; store at room temperature • Compatible—use on nitrocellulose and PVDF membranes, whether still wet or already dry; works with practically any blocking buffer, enzyme conjugate and chemiluminescent substrate • Cost effective—save valuable time and samples; strip blots effectively the first time • Robust yet gentle—transferred proteins remain viable; strip the same blot up to five times • Flexible—strip and reprobe to optimize antibody concentrations or to detect a new antigen with different antibodies
Restore PLUS Buffer is an alternative formulation of the original Thermo Scientific Restore Western Blot Stripping Buffer. Restore PLUS Stripping Buffer was designed for use with antibodies that are difficult to remove from western blots and require longer incubation times or incubation temperatures greater than 22°C with gentler formulations. High-affinity antibodies can be quickly and effectively stripped from western blots at room temperature without removing transferred proteins, thereby allowing multiple reprobes of the target.
Protocol Summary: • Wash blot to remove chemiluminescent substrate • Incubate blot in Restore Western Blot Stripping Buffer for 5 to 15 minutes at room temperature (or incubate at 37°C for high affinity antibodies) • Remove blot and wash in Wash Buffer • Block membrane • Test for sufficient removal of antibodies • Perform next immunoblot experiment
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeWestern Blot Stripping Buffer
Detection MethodChemiluminescence
FormLiquid
For Use With (Application)Western Blot
Membrane CompatibilityNitrocellulose, PVDF
Product LineRestore™
Quantity30 mL
Unit Size30 mL
Contents & Storage
Upon receipt store at room temperature. Product shipped at ambient temperature.
Frequently asked questions (FAQs)
How many times can I strip my blot using Restore or Restore Plus Western Blot Stripping Buffers ?
The stability of the attached (transferred and bound) protein will determine the number of times the membrane can be successfully re-probed after stripping. The protein may withstand stripping as many as four times or as few as one time.
Can I strip membranes with Restore or Restore Plus Western Blot Stripping Buffer and reprobe for subsequent detection with fluorescent probes?
No. Although the fluorescent antibodies, like other antibodies, can be stripped with Restore Buffers, stripped membranes typically produce unacceptable background for subsequent fluorescent detection methods. We recommend Restore Fluorescent Western Blot Stripping Buffer, Cat. No. 62299 (20 mL) and Cat. No. 62300 (100 mL).
Note: Restore Fluorescent Western Blot Stripping Buffer is for use with low-fluorescence PVDF membrane (Cat. No. 22860) only.
Will Restore and Restore Plus Western Blot Stripping Buffers remove biotin-bound avidin, streptavidin or NeutrAvidin Protein conjugates?
No. Restore and Restore Plus Buffers are not stringent enough to break the avidin-biotin interaction. A buffer capable of breaking this interaction would likely also damage the target protein, making it undetectable. However it may be possible to strip the complexed biotinylated primary antibody and streptavidin-HRP secondary antibody from the target protein on the membrane. Please also see Tech Tip: Strip and Reprobe Western Blots (https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/strip-reprobe-western-blots-tech-tip.pdf).
Will the Restore and Restore Plus Western Blot Stripping Buffers remove precipitating substrates?
No. The antibodies are removed but the substrate leaves a permanent precipitate on the membrane that cannot be removed. Restore and Restore Plus Buffers are designed for procedures using chemiluminescent substrates. Please note that this is not compatible with fluorescence supstrates as it will result in increased background. For fluorescent substrates please use our Restore Fluorescent Western Blot Stripping Buffer. Please also see Tech Tip: Strip and Reprobe Western Blots (https://assets.thermofisher.com/TFS-Assets/BID/Technical-Notes/strip-reprobe-western-blots-tech-tip.pdf).