Update profile
    Change password
    Sign out

Our content has moved! Check out our protein biology page on lifetechnologies.com  Go now >

Desthiobiotin Product Selection Guide

Learn about desthiobiotin and other elutable or reversible avidin-biotin affinity systems.

Desthiobiotin Reagent GuideDesthiobiotin is a modified form of biotin that binds less tightly to avidin and streptavidin than biotin while still providing excellent specificity in affinity purification methods. Unlike biomolecules that are labeled with biotin, proteins and other targets that are labeled with desthiobiotin can be eluted without harsh, denaturing conditions. This guide describes available desthiobiotin labeling reagents and demonstrates desthiobiotinylation as an alternative to biotinylation for applications requiring gentle elution and recovery of the labeled target. Other reversible avidin-biotin binding methods are also mentioned.

Page Contents:

Learn more...

Protein Methods Library Home


View products...

Desthiobiotinylation Reagents

All Biotin Labeling Reagents

Desthiobiotin vs. biotin

Limitations of biotin for affinity purification

Because of its especially high affinity and binding specificity, the biotin-avidin (or biotin-streptavidin) interaction has been used for several decades as the basis of many laboratory research techniques to label, detect and purify biomolecules. However, one limitation of the biotin-avidin interaction in purification applications is that it is essentially irreversible under physiological conditions. For example, once the biotinylated proteins in a sample are successfully captured by streptavidin agarose or magnetic beads, they cannot be dissociated (eluted and recovered) without using harsh conditions (boiling, extreme pH, or denaturants). These conditions tend to denature the proteins of interest or cause artifacts in downstream applications, such as gel electrophoresis.


Desthiobiotin structure and properties

To overcome this limitation, new reagents and assays based on desthiobiotin, a non-sulfur containing biotin analog, are being more widely developed and utilized. Desthiobiotin binds less tightly to avidin and streptavidin than biotin (Kd = 10^-11 M versus Kd = 10^-15 M, respectively) but nonetheless provides a high level of specificity.

Comparison of the chemical structures of biotin and desthiobiotin
Comparison of the chemical structures of biotin and desthiobiotin.
View products...

Desthiobiotinylation Reagents

All Biotin Labeling Reagents

Unlike biotinylated proteins, desthiobiotinylated bait proteins and their interacting partners can be readily and specifically eluted under mild conditions when captured on streptavidin by using a biotin elution buffer. The soft release characteristic of desthiobiotin minimizes the isolation of naturally biotinylated molecules that can interfere with results and also eliminates the use of harsh elution conditions which can disassociate complexes and/or damage the target protein or cell. This technique is ideal when using native or recombinant proteins that are not expressed with a fusion tag and when you want to isolate captured proteins under native conditions such as targeting intact cells or cell surface proteins.

Desthiobiotin provides more effect and gentler elution than biotin for streptavidin-based affinity purification
Elution comparison between desthiobiotin and biotin in pull-down experiments using streptavidin magnetic beads. Purified IgG was labeled with equivalent desthiobiotin or biotin reagents, spiked into A431 cell lysate, and then purified using streptavidin magnetic beads. Different fractions from each pull-down experiment were electrophoresed and the IgG was detected by Western blot. S = starting labeled IgG samples; FT = flow-through; E1-E2 = elution fractions (desthiobiotin pull-down eluted with 4mM biotin; biotin pull-down eluted with 0.5% formic acid, 30% acetonitrile); BB1-BB2 = bead boil fractions. As the red arrows indicate, desthiobiotinylated IgG elutes easily by gentle, competitive displacement with 4mM biotin, while biotinylated IgG elutes effectively only upon boiling.


View products...

Avidin, Streptavidin and other
Biotin-Binding Affinity Resins

Streptavidin Magnetic Beads



Menu of desthiobiotin products

Menu of EZ-Link Desthiobiotin Reagents.
Products Activation Labeling application Learn
NHS-Desthiobiotin NHS Primary amines
(lysines) in proteins
Sulfo-NHS-LC-Desthiobiotin Sulfo-NHS Primary amines
(lysines) in proteins
Amine-PEG4-Desthiobiotin Amine Carboxyl groups
(via EDC crosslinker)
Hydrazide-PEG4-Desthiobiotin Hydrazide Carbonyl groups
(oxidized carbohydrates in glycoproteins)
Phosphine-PEG4-Dethiobiotin Phosphine Azide groups
(created by metabolic labeling)
Staudinger ligation


Menu of assays and kits that use desthiobiotin.
Products Desthiobiotin reagent Application
Desthiobiotinylation and Pull-Down Kit Sulfo-NHS-LC-Desthiobiotin Label a purified protein and use it to affinity purify interaction complexes in which it participates
RNA 3' End Desthiobiotinylation Kit Desthiobiotinylated Cytidine Bisphosphate Enzymatically label an RNA oligo for use in Magnetic RNA-Protein Pull-Down Kit
Active Serine Hydrolase Probes Desthiobiotin-fluorophosphonate Covalently label the active site of serine hydrolases to enable their selective enrichment
GTPase Enrichment Kit Desthiobiotin-GTP Covalently label the active site of GTPases and the GTPase subunits of G-protein coupled receptors to enable their selective enrichment
Kinase Enrichment Kits Desthiobiotin-ATP;
Covalently label the active site of ATPases, including chaperones and metabolic enzymes, to enable their selective enrichment


Other reversible avidin-biotin strategies

Modified avidin

Elutable avidin-biotin systems have been created by modifying the avidin protein so that it has lower affinity towards biotin. Monomeric Avidin Agarose, which consists of avidin that has been immobilized and maintained primarily in monomeric form, and CaptAvidin™ Protein, which has a nitrated tyrosine group in its biotin-binding site, are examples of this sort of strategy.

View products...

Monomeric Avidin Agarose

CaptAvidin Protein
(Life Technologies website)

Cleavable biotin reagents

Another approach is to use biotinylation reagents that contain a disulfide bond (-S-S-) or other easily cleavable group in their spacer arms; Sulfo-NHS-SS-Biotin is an example of such a reagent. A protein labeled with this reagent can be recovered from its bound state with streptavidin resin by reduction (cleavage) of the disulfide bond in spacer arm between the biotin and protein; in this case, the biotin group is left bound to the streptavidin and the protein is recovered without a biotin label. However, native disulfide bonds in the protein of interest also may be cleaved with this method. Sulfo-NHS-SS-Biotin is used as the basis for isolation of cell surface proteins in the Pierce Cell Surface Isolation Kit.

View products...

Cell Surface Protein Isolation Kit






Sulfo-SBED Biotin Label Transfer Reagent


Iminobiotin is a cyclic guanidino analog of biotin that exhibits considerably weaker binding to avidin. This form of biotin has limited utility in most practical applications. Its primary use is as an affinity resin for purifying functional streptavidin or avidin proteins.

View products...

Iminobiotin Agarose


  1. Hirsch JD, et al. (2002). Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation. Anal Biochem. 308:343-57.
  2. Hofmann K, et al. (1984). Synthesis of biotinylated and dethiobiotinylated insulins. Biochem. 23:2547-53.
  3. Hofmann K, et al. (1982). Avidin binding of carboxyl-substituted bihotin and analogues. Biochem. 21:978-84.

Browse all Desthiobiotin products

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

Recommend this page

Follow Us
Email Sign-up  Email Announcements


Antibodies  |   Molecular Biology   |   Cell Biology   |  Thermo Scientific  |   * Trademarks  |   Privacy Statement
© 2015 Thermo Fisher Scientific Inc.   |   3747 N Meridian Rd, Rockford, IL USA 61101   |   1-800-874-3723  or  815-968-0747