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If I want to strip and re-probe a nitrocellulose membrane, what procedure do you recommend? Product FAQ

Answer

Thermo Fisher Scientific sells 4 ready-to-use buffers specifically intended for stripping Western blots prior to re-probing. Three of them can be used with both nitocellulose and PVDF membranes. We have Restore Western Blot Stripping Buffer (Cat. No. 21059), a trial size of this product (Cat. No. 21062), a product called Restore PLUS (Cat. No. 46428), which is formulated to strip off antibodies that are difficult to remove, and Restore Fluorescent Western Blot Stripping Buffer (Cat. No. 62299). The latter is used to strip fluorophore-labeled antibodies from PVDF membranes only.

Answer Id:: E5217

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Products for Western Blotting Product Literature

Assessment of inactivated human rabies vaccines: biochemical characterization and genetic identification of virus strains. Citations & References

  • Authors: Finke S, Karger A, Freuling C, Müller T,
  • Journal: Vaccine
  • PubMed ID: 22469862
Catalog #

Physiological and proteomic approaches to address the active role of ozone in kiwifruit post-harvest ripening. Citations & References

  • Authors: Minas IS, Tanou G, Belghazi M, Job D, Manganaris GA, Molassiotis A, Vasilakakis M,
  • Journal: J Exp Bot
  • PubMed ID: 22268155
Catalog #

Article: Biological validation of a novel process and product for quantitating western blots Product Literature

Can I use fluorescent secondary antibodies for western blots? Product FAQ

Answer

Yes, as long as the instrument used to analyze the blots provides the appropriate excitation wavelengths and emission filter sets to detect fluorescence of the selected dye(s). The limits of detection are dependent upon the amount of protein/peptide per spot/band, using the correct excitation and emission settings for the dye-conjugated antibody, and the sensitivity of the camera.
Another alternative to dye-conjugated secondary antibodies are the quantum dot-conjugated antibodies provided in our WesternDot products. For more information: https://www.thermofisher.com/us/en/home/life-science/protein-biology/protein-assays-analysis/western-blotting/detect-proteins-western-blot/western-blot-detection-reagents/fluorescent-western-blot-detection/westerndot-fluorescent-immunodetection-kits.html

Answer Id:: E18039

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Has the BenchPro 4100 Card Processing Station been discontinued? Product FAQ

Answer

Yes, the BenchPro 4100 Card Processing Station has been discontinued as of December 31, 2014. The current BenchPro 4100 Western Cards (https://www.thermofisher.com/order/catalog/product/WP1001) and Reagent Vials (https://www.thermofisher.com/order/catalog/product/WP3001) will continue to be available for purchase. We offer the new iBind Western Device (http://www.thermofisher.com/us/en/home/life-science/protein-expression-and-analysis/western-blotting/western-blot-detection/ibind-western-system.html) as an alternative system for automated western blot processing.

Answer Id:: E10882

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Highly specific confirmatory western blot test for African swine fever virus antibody detection using the recombinant virus protein p54. Citations & References

  • Authors: Alcaraz C; Rodriguez F; Oviedo JM; Eiras A; De Diego M; Alonso C; Escribano JM
  • Journal: Journal of Virological Methods
Catalog #

What is the difference between the Restore and Restore Plus Western Blot Stripping Buffers? Product FAQ

Answer

Restore Western Blot Stripping Buffer gently but effectively removes the primary and secondary antibodies from the membrane to allow re-probing on the same membrane. The buffer is formulated to work for a wide variety of interactions, but there are some high affinity antigen-antibody interactions that require more stringent stripping conditions. Restore Plus Western Blot Stripping Buffer was developed for these difficult to strip interactions.

Answer Id:: E8463

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What Western blot stripping buffers do you offer? Product FAQ

Answer

There are two types of stripping buffers, each available in two different sizes:

• Restore Western Blot Stripping Buffer: Cat. No. 21059 (500 mL), Cat. No. 21063 (5 L) and Cat. No. 21062 (30 mL)
• Restore Plus Western Blot Stripping Buffer: Cat. No. 46430 (500 mL) and Cat. No. 46428 (30 mL)

Answer Id:: E8462

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Have the Restore and Restore Plus Western Blot Stripping Buffers been tested for my primary or secondary antibody? Product FAQ

Answer

No. Every binding interaction is slightly different in terms of mode (i.e., actual amino acids or functional groups that interact) and affinity (strength of binding under a given set of buffer conditions). First try Restore Western Blot Stripping Buffer for 15 minutes at room temperature. If antibody removal is incomplete, optimize the stripping conditions by increasing the time and temperature. If this fails to completely strip the antibodies, then switch to the more stringent Restore Plus Western Blot Stripping Buffer.

Answer Id:: E8464

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Will Restore and Restore Plus Western Blot Stripping Buffers remove biotin-bound avidin, streptavidin or NeutrAvidin Protein conjugates? Product FAQ

Answer

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 # 23 Strip and Reprobe Western Blots (https://tools.thermofisher.com/content/sfs/brochures/TR0023-strip-reprobe-blots.pdf).

Answer Id:: E8469

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Tech Tip: Strip and reprobe Western blots Product Literature

Tech Tip: Strip and reprobe Western blots Product Literature

What applications are appropriate for your Anti-Xpress antibody, Anti-V5 antibody, Anti-HisG antibody, and Anti-myc antibody? Product FAQ

Answer

ELISA, Western blot, Immunoprecipitation, Immunofluorescence, and Immunohistochemistry protocols have all been used successfully with our epitope tag antibodies. Please consult the product manuals for specific protocols and dilutions.

Answer Id:: E3641

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