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EZ-Link Hydrazide-PEG4-Desthiobiotin

Label glycoproteins for affinity purification with streptavidin beads.

EZ-Link Hydrazide-PEG4-Desthiobiotin

Thermo Scientific EZ-Link Hydrazide-PEG4-Desthiobiotin is a pegylated, carbonyl-reactive labeling reagent whose biotin-like group is elutable from streptavidin, enabling its use for cell surface glycoprotein labeling and purification.

This desthiobiotin variant of biotin is activated to label periodate-oxidized carbohydrates of macromolecules. The hydrazide group conjugates to carbonyls (aldehydes and ketones), such as those created from sialic acid and other sugar components of glycoprotein polysaccharides upon treatment with 1 to 10mM sodium periodate (NaIO4). The desthiobiotin tag binds to streptavidin and other biotin-binding proteins with high specificity yet readily elutes with mild conditions (i.e., by competitive displacement with regular, free biotin). As such, this reagent is a useful alternative to Hydrazide-PEG4-Biotin (Part No. 21360) for avidin-biotin techniques in which nondenaturing elution of the labeled proteins is desired. The hydrophilic, 4-unit polyethylene glycol (PEG) spacer arm enhances solubility and reduces aggregation of labeled proteins stored in solution. The PEG segment also adds length and flexibility to the spacer arm, minimizing steric hindrance involved with binding to avidin molecules. The No-Weigh Format (5 x 1mg resealable vials) eliminates the difficulties associated with weighing small quantities of reagent and maximizes protection of unused reagent from degradation.


  • Desthiobiotin – analog of biotin allows easy elution from streptavidin, an ideal feature for affinity purification applications
  • Glycoprotein labeling – tag glycosylated proteins at sialic acid residues to enable high recovery in pull-down assays with streptavidin beads
  • Cell surface labeling – tag and isolate cell surface glycoproteins; reagent does not permeate membranes of whole cells
  • Aldehyde-reactive – reacts with aldehydes formed by periodate-oxidation of sugar groups
  • Hydrazide-activated – perform reactions at pH 4 to 6 in buffers such as sodium acetate
  • Pegylated – spacer arm contains a hydrophilic, 4-unit, polyethylene glycol (PEG) group
  • Enhances solubility – pegylation imparts water solubility to the biotinylated molecule, helping to prevent aggregation of biotinylated antibodies stored in solution

Product Details:

Chemical structure of NHS-SS-Biotin
Chemical structure of Hydrazide-PEG4-Desthiobiotin. This reagent labels glycoproteins and other molecules having carbohydrates that can be oxidized with periodate to expose aldehydes. For more information, see our review of Hydrazide Reaction Chemistry.

Properties of Hydrazide-PEG4-Desthiobiotin. We manufacture our reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.
Molecular formula C21H41N5O7
Molecular weight 475.58
Spacer arm length 31.2 angstroms
Mass added to target 457.26 daltons
Storage conditions 4°C; -20°C as stock solution
Reactive groups Hydrazide, reacts with aldehydes at pH 4 to 6

Desthiobiotin vs. Biotin:

Desthiobiotin is a single-ring, sulfur-free analog of biotin that binds to streptavidin with nearly equal specificity but less affinity than biotin (1/Kd = 10^11 vs. 10^15M, respectively)[1-2]. Consequently, desthiobiotinylated bait proteins and their interacting partners can be eluted readily and specifically from streptavidin affinity resin using mild conditions based on competitive displacement with free biotin. For pull-down assay experiments with biological samples, this soft-release characteristic of desthiobiotin also helps to minimize co-purification of endogenous biotinylated molecules, which remain bound to streptavidin upon elution of the target protein complexes with free biotin. The modified avidin-biotin affinity system also eliminates the need to use harsh elution conditions that might disassociate complexes and/or damage the target protein or cell. Desthiobiotin-based techniques are ideal when using native or recombinant proteins that are not expressed with a fusion tag and when isolating captured proteins under native conditions, such as targeting intact cells or cell surface proteins.

Labeling with Hydrazide Reagents:

Hydrazides and alkoxyamines are two types of carbonyl-reactive groups. Hydrazides (–NH-NH2) react specifically with aldehyde groups in slightly acidic conditions to form hydrazone linkages; these can be further reduced to stable secondary amine bonds using sodium cyanoborohydride (Part No. 44892). The reaction is more efficient in the presence of aniline (Part No. 88944). Alternatively, hydrazides can be conjugated to carboxylic acids using EDC carbodiimide chemistry.

Reactive aldehyde groups can be generated in glycoproteins and other polysaccharide compounds by oxidation of constituent sugar diols using sodium periodiate (Part No. 20504). Sialic acid residues are common components of protein glycosylation and are easily converted to aldehydes with 1mM NaIO4.

Cited References:

  1. Hirsch, J., et al. (2002). Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection, and isolation. Analytical Biochemistry 308: 343-357.
  2. Hofmann, K., et al. (1982). Avidin binding of carboxyl-substituted biotin and analogues. Biochemistry 21: 978-984.

Related Resources:

Review of Biotinylation Methods and Applications
Chemistry of Crosslinking (and Labeling Reagents)

Related Products:

Desthiobiotin Product Selection Guide
All Biotinylation Reagents
Avidin, Streptavidin, NeutrAvidin Affinity Resins

Ordering Information
Product # Description Pkg. Size Instructions MSDS CofA Price
16130 EZ-Link Hydrazide-PEG4-Desthiobiotin, No-Weigh Format
Formulation: Hydrazide-activated desthiobiotin with a tetraethyleneglycol spacer
Sufficient For: 5 uses to label a total of 2mg of IgG using typical conditions
5 x 1mg Product Instructions for product #16130 EZ-Link Hydrazide-PEG4-Desthiobiotin, No-Weigh Format MSDS for product #16130 EZ-Link Hydrazide-PEG4-Desthiobiotin, No-Weigh Format Certificate of Analysis for product #16130 EZ-Link Hydrazide-PEG4-Desthiobiotin, No-Weigh Format $405.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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