Manual / Product Insert

User Guide: Biotin Chromogenic Detection Kit

Version: MAN0012660 Rev 9 (Sep 2015 )

Citations & References

Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation

  • Authors: Hirsch, J., et al.
  • Journal: Analytical Biochemistry (2002) 308:343-357
Catalog #
  • 16129
  • 16138
  • 16130(Discontinued)
  • 16131(Discontinued)
  • 16133(Discontinued)
  • 16136(Discontinued)

Citations & References

Biotin-pyrene conjugates with poly(ethylene glycol) spacers are convenient fluorescent probes for avidin and streptavidin.

  • Authors: Marek M, Kaiser K, Gruber HJ
  • Journal: Bioconjug Chem (1997) 8:560-566
  • PubMed ID: 9258456
Catalog #
  • B1370
  • P130
  • A50(Discontinued)
  • P1903MP(Discontinued)
  • P32

Citations & References

Application of biotin-4-fluorescein in homogeneous fluorescence assays for avidin, streptavidin, and biotin or biotin derivatives.

  • Authors: Ebner A, Marek M, Kaiser K, Kada G, Hahn CD, Lackner B, Gruber HJ,
  • Journal: Methods Mol Biol (2008) 418:73-88
  • PubMed ID: 18287651
Catalog #

Product FAQ

What protocol is recommended for removal of residual biotin, avidin or streptavidin activity from tissue sections?

Answer

For removal of residual biotin: Incubate sections with the avidin or streptavidin solutions from the Avidin/Biotin Blocking kit (Cat. No. 00-4303) or the Endogenous Biotin-Blocking Kit (Cat. No. E21390), respectively. Then wash, as directed. Incubate with the appropriate d-biotin solutions included in the kits. Wash thoroughly again.

For removal of residual avidin or streptavidin: Incubate sections with the appropriate d-diotin solution from either the Avidin/Biotin Blocking kit (Cat. No. 00-4303) or the Endogenous Biotin-Blocking Kit (Cat. No. E21390). Wash thoroughly.

Blocking residual biotin, avidin or streptavidin activity should be performed immediately after your normal blocking step, but before your primary antibody incubation. This blocks endogenous biotin exposed by the antigen retrieval procedure as well as any endogenous biotin that may be present in the blocking solution.

Answer Id: E4912x

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Citations & References

Studies on the biotin-binding site of avidin. Minimized fragments that bind biotin.

  • Authors: Hiller Y, Bayer EA, Wilchek M
  • Journal: Biochem J (1991) 278 ( Pt 2):573-585
  • PubMed ID: 1898347

Citations & References

Biotin-avidin immobilization of platelet glycoproteins (BAIPG): a new capture assay for the detection of anti-platelet antibodies.

  • Authors: Cordiano I, Steffan A, Randi ML, Pradella P, Girolami A, Fabris F
  • Journal: J Immunol Methods (1995) 178:121-130
  • PubMed ID: 7829861

Product FAQ

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

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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Citations & References

A monovalent streptavidin with a single femtomolar biotin binding site.

  • Authors: Howarth M, Chinnapen DJ, Gerrow K, Dorrestein PC, Grandy MR, Kelleher NL, El-Husseini A, Ting AY
  • Journal: Nat Methods (2006) 3:267-273
  • PubMed ID: 16554831

Citations & References

Endogenous biotin in rat brain: implications for false-positive results with avidin-biotin and streptavidin-biotin techniques.

  • Authors: McKay BE, Molineux ML, Turner RW,
  • Journal: Methods Mol Biol (2008) 418:111-128
  • PubMed ID: 18287654

Product FAQ

What is the affinity of streptavidin for biotin in the Thermo Scientific Streptavidin Plus Ultralink Resin kit?

Answer

The binding affinity of streptavidin is similar to that of avidin, which is Ka = 10^15/M.

Answer Id: E8255

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Citations & References

Evaluation of biotin-dye conjugates for use in an HPLC assay to assess relative binding of biotin derivatives with avidin and streptavidin.

  • Authors: Wilbur DS, Pathare PM, Hamlin DK, Frownfelter MB, Kegley BB, Leung WY, Gee KR
  • Journal: Bioconjug Chem (2000) 11:584-598
  • PubMed ID: 10898581
Catalog #

Citations & References

Accurate measurement of avidin and streptavidin in crude biofluids with a new, optimized biotin-fluorescein conjugate.

  • Authors: Kada G, Falk H, Gruber HJ
  • Journal: Biochim Biophys Acta (1999) 1427:33-43
  • PubMed ID: 10082985
Catalog #

Citations & References

Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation.

  • Authors: Hirsch JD, Eslamizar L, Filanoski BJ, Malekzadeh N, Haugland RP, Beechem JM, Haugland RP
  • Journal: Anal Biochem (2002) 308:343-357
  • PubMed ID: 12419349
Catalog #