Trusted by Leading Research and Pharma Institutions

Enzymatic Biotinylation Service

Precision protein labeling powered by E. coli biotin ligase (BirA) and AviTag technology. Achieve homogeneous, site-specific biotinylation with our validated enzymatic platform for complete positional specificity.

100% Site-Specific

MS Verified

Minimal Tag Footprint

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Trusted by leading research and pharmaceutical institutions

Harvard

Pfizer

MIT

Roche

Stanford

Novartis

Why Choose Enzymatic Labeling

Homogeneous single-site modification

Near-complete biotinylation efficiency

Mild physiological conditions

15-amino acid AviTag minimal footprint

BirA Biotin Ligase

E. coli enzyme for site-specific modification

Quality Guaranteed

Mass spec verification on every batch

Flexible Placement

N-terminus, C-terminus, or internal loops

Efficiency

95%+

Service Overview

Why Choose Enzymatic Biotinylation

Our platform combines the precision of enzymatic catalysis with the power of the AviTag system for unmatched labeling consistency.

100% Site-Specific

Enzymatic biotinylation targets a single lysine residue within the AviTag sequence, producing a homogeneous product with complete positional specificity that chemical methods cannot achieve.

Single modification site guaranteed
Complete batch-to-batch consistency
No heterogeneous mixtures

High Efficiency

BirA enzyme achieves near-complete biotinylation under mild reaction conditions, preserving protein function and activity throughout the process.

Typically achieves 100% modification
Mild pH 7.5-8.0 conditions
No harsh chemicals or reagents

Verified Purity

Every batch undergoes mass spectrometry verification to confirm complete biotinylation.

Minimal Tag Footprint

The AviTag is only 15 amino acids - far smaller than traditional affinity tags.

Mild Conditions

Reaction occurs under physiological temperature and pH, preserving protein structure.

Reversible Capture

Desthiobiotin analogs enable reversible streptavidin binding with high affinity.

Technology Platform

Our Enzymatic Biotinylation Technology

How BirA Biotinylation Works

Biotin Activation

BirA catalyzes the formation of biotinyl-5'-AMP intermediate in the presence of biotin and ATP.

Site Recognition

The enzyme specifically recognizes the 15-amino acid AviTag sequence (GLNDIFEAQKIEWHE).

Covalent Attachment

Biotin is covalently attached to the lysine residue within the AviTag, forming a stable amide bond.

Quality Verification

Mass spectrometry confirms complete biotinylation and product homogeneity.

AviTag Sequence:

G L N D I F E A Q K I E W H E

K = Biotin attachment site

Lysine (K) at position 10 is the specific biotinylation site

Amino Acids

Modification Site

Single Site Modification

Unlike chemical biotinylation which produces heterogeneous mixtures, enzymatic labeling ensures 100% site uniformity.

Homogeneous Reproducible

Mild Conditions

Reaction occurs under physiological pH and temperature, preserving sensitive protein structures and activities.

pH 7.5-8.0 30C Optimal

Reversible Capture

Desthiobiotin analogs enable reversible streptavidin binding while maintaining the same high affinity.

Reusable Cost-Effective

Specifications

Service Specifications

Comprehensive specifications to meet your research requirements.

Parameter	Specification
Tag Sequence	GLNDIFEAQKIEWHE (AviTag, 15 amino acids)
Biotinylation Efficiency	Greater than 95% (verified by MS, typical: 100%)
Reaction Temperature	30C (optimal), also at RT or 4C
Reaction Time	30 min - 2 hours (depending on conditions)
Buffer Conditions	PBS, Tris-HEPES pH 7.5-8.0, with 150mM NaCl
Tag Placement	N-terminus, C-terminus, or internal loops
Protein Requirements	10-50 micromolar protein concentration recommended
Quality Control	SDS-PAGE, Gel shift assay, Mass spectrometry
Purification	Ni-NTA reverse purification, SEC cleanup

100% Biotinylation

MS Verified

Endotoxin-Free

Batch Consistency

Workflow

Streamlined Process from Design to Delivery

Our proven workflow ensures quality and efficiency at every stage.

Consultation

Project review and AviTag placement optimization

Construct

Gene synthesis with AviTag insertion

Expression

Recombinant protein production

Biotinylation

BirA enzymatic modification

QC & Delivery

MS verification and product shipment

Applications

Diverse Applications Across Biotechnology

Enzymatically biotinylated proteins support research and development in multiple fields.

Protein Purification

Biotinylated proteins can be efficiently captured using streptavidin-coated beads or columns, providing a powerful alternative to traditional affinity tags like His-tag or GST with superior binding capacity.

High binding capacity with streptavidin matrices
Stable binding under various conditions
Compatible with denaturing purification
Can be eluted under mild conditions

Strong

Streptavidin-Biotin Binding

Detection and Imaging

Use biotinylated proteins with fluorescently-conjugated streptavidin for highly sensitive detection in Western blot, ELISA, flow cytometry, and live-cell imaging applications.

Western blot with streptavidin-HRP
ELISA with streptavidin detection
Flow cytometry analysis
Live-cell imaging with fluorophore conjugates

Sensitive

Detection Methods

Protein-Protein Interactions

Biotinylated proteins enable precise study of protein interactions through pull-down assays, surface plasmon resonance (SPR), and molecular bridging experiments.

Co-immunoprecipitation (Co-IP)
Surface plasmon resonance (SPR)
Protein complex isolation
Molecular bridging assays

Precise

Interaction Studies

Surface Immobilization

Oriented immobilization of biotinylated proteins on streptavidin-coated surfaces ensures uniform presentation and maximum accessibility for binding assays.

Biosensor chip functionalization
Microarray spot preparation
Nanoparticle conjugation
Single-molecule studies

Oriented

Surface Immobilization

Testimonials

What Our Clients Say

Trusted by researchers worldwide for quality and reliability.

The biotinylation efficiency exceeded our expectations. Complete modification at a single site simplified our downstream applications significantly.

Senior Scientist

Biotechnology Company

We have been using this service for our MHC tetramer production. The homogeneous biotinylation ensures consistent staining in flow cytometry experiments.

Principal Investigator

Academic Research Institution

The technical support team helped us optimize AviTag placement for a challenging membrane protein. Excellent service and reproducible results every time.

Research Director

Pharmaceutical Company

Scientific Literature

Scientific Foundation

Our platform is backed by peer-reviewed research.

412 Citations

Site-specific biotinylation of purified proteins using BirA

Fairhead M, Howarth M. Methods in Molecular Biology (Springer), 2015.

Comprehensive protocols for AviTag insertion, BirA purification, and gel-shift analysis to quantify biotinylation extent.

View DOI

2024

A one-process production of completely biotinylated proteins in a T7 expression system

Kawashima T, Nakamura M, Sakono M. Biotechnology and Applied Biochemistry, 2024.

Establishes an in vivo method for complete biotinylation using single plasmid co-expressing BirA and target proteins in E. coli.

View DOI

28 Citations

Off-the-shelf proximity biotinylation using ProtA-TurboID

Santos-Barriopedro I, van Mierlo G, Vermeulen M. Nature Protocols, 2023.

Protocol for using Protein A-TurboID enzyme for antibody-based proximity biotinylation in various cell types.

View DOI

1350 Citations

Efficient proximity labeling in living cells and organisms with TurboID

Branon TC, Bosch JA, Sanchez AD, et al. Nature Biotechnology, 2018.

Engineering of TurboID and miniTurbo enzymes for fast, efficient proximity labeling with non-toxic biotin.

View DOI

2024

Exploring Options for Proximity-Dependent Biotinylation Experiments: Comparative Analysis of Labeling Enzymes and Affinity Purification Resins

Schreiber KJ, Kadijk E, Youn JY. Journal of Proteome Research (ACS), 2024.

Systematic comparison of miniTurbo and APEX2 enzymes with different streptavidin resins for proximity biotinylation.

View DOI

FAQ

Frequently Asked Questions

Find answers to common questions about our service.

What is the AviTag sequence?

The AviTag is a 15-amino acid peptide with the sequence GLNDIFEAQKIEWHE. It is specifically recognized by E. coli biotin ligase (BirA), which covalently attaches biotin to the lysine residue (K) at position 10. The AviTag is significantly smaller than traditional affinity tags, minimizing its impact on protein folding and function.

How do you verify 100% biotinylation efficiency?

We use multiple verification methods: (1) Streptavidin gel shift assay - the biotinylated protein shows a clear mass shift upon streptavidin binding on SDS-PAGE; (2) High-resolution mass spectrometry for absolute molecular weight confirmation; (3) Quantification of unmodified versus modified protein. Every batch comes with a Certificate of Analysis and MS verification report.

Where should I place the AviTag on my protein?

The AviTag can be placed at the N-terminus, C-terminus, or within accessible loops of your target protein. We recommend adding 2-3 flexible residues (Gly-Ser) on either side of the AviTag to insulate it from the folded protein structure. Our scientific team can perform structural modeling to identify optimal placement that ensures accessibility to BirA while preserving protein function.

What is the difference between enzymatic and chemical biotinylation?

Enzymatic biotinylation using BirA produces homogeneous, site-specific modification at a single lysine within the AviTag sequence. Chemical biotinylation (using NHS-biotin or other reagents) modifies all accessible primary amines on the protein surface, resulting in heterogeneous products with variable biotinylation stoichiometry. Enzymatic labeling ensures consistent batch-to-batch results and preserves protein function.

Will the BirA enzyme remain in the final product?

No. Our workflow includes purification steps (such as reverse Ni-NTA affinity chromatography for His-tagged BirA, and/or size-exclusion chromatography) to ensure complete removal of the BirA enzyme, unreacted biotin, and ATP from the final biotinylated protein product.

Can you work with proteins that already have an AviTag?

Yes. If you already have an AviTag-fusion construct, we can proceed directly to the biotinylation reaction step. Simply provide us with your AviTag-fusion protein, and we will perform the BirA-mediated biotinylation with full quality verification. We can also help you clone your gene into our AviTag expression vectors if needed.

What are the storage conditions for biotinylated proteins?

Biotinylated proteins are typically provided as lyophilized powder or in solution. For long-term storage, we recommend -80C. The biotin-protein bond is highly stable and can withstand multiple freeze-thaw cycles. Avoid repeated freezing if possible; aliquoting is recommended. When stored properly, biotinylated proteins remain functional for extended periods.

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Enzymatic Biotinylation Service

Why Choose Enzymatic Labeling

BirA Biotin Ligase

Quality Guaranteed

Flexible Placement

Why Choose Enzymatic Biotinylation

100% Site-Specific

High Efficiency

Verified Purity

Minimal Tag Footprint

Mild Conditions

Reversible Capture

Ready to Get Started?

Our Enzymatic Biotinylation Technology

How BirA Biotinylation Works

Biotin Activation

Site Recognition

Covalent Attachment

Quality Verification

Single Site Modification

Mild Conditions

Reversible Capture

Service Specifications

Streamlined Process from Design to Delivery

Consultation

Construct

Expression

Biotinylation

QC & Delivery

Diverse Applications Across Biotechnology

Protein Purification

Detection and Imaging

Protein-Protein Interactions

Surface Immobilization

What Our Clients Say

Scientific Foundation

Site-specific biotinylation of purified proteins using BirA

A one-process production of completely biotinylated proteins in a T7 expression system

Off-the-shelf proximity biotinylation using ProtA-TurboID

Efficient proximity labeling in living cells and organisms with TurboID

Exploring Options for Proximity-Dependent Biotinylation Experiments: Comparative Analysis of Labeling Enzymes and Affinity Purification Resins

Frequently Asked Questions

Ready to Start Your Project?

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