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Biotinylation Reagents

Biotinylation Reagents & Streptavidin Systems

Introduction

Biotinylation is a powerful technique for labeling proteins and nucleic acids. APExBIO offers a complete ecosystem of biotin reagents (NHS-ester, Maleimide, PEG-linkers) and high-affinity Streptavidin conjugates (HRP, Fluorophores, Magnetic Beads) for high-sensitivity detection and purification.

Overview

What is Biotinylation?

Biotinylation is the process of covalently attaching Biotin (Vitamin H) to a target molecule. This leverages the Biotin-Streptavidin interaction, which is the strongest non-covalent biological bond (Kd ≈ 10-14M).

Application

  • Protein Enrichment: Using Streptavidin magnetic beads for pull-down assays.
  • Surface Labeling: Identifying membrane proteins via non-permeable biotin tags.
  • Signal Amplification: Enhancing Western Blot and ELISA sensitivity using Streptavidin-HRP.
  • Proximity Labeling: Mapping protein-protein interactions (PPI) in live cells.

Workflow

Biotin-Streptavidin System

Choosing the Right Biotinylation Reagent

You can choose biotin tags based on their reactivity and solubility:

Selection

Selected By Features Application Proudct
Solubility & Membrane Permeability
Water Soluble Membrane-Impermeable
Features a sulfonate group, ideal for aqueous reactions. Gold standard for Cell Surface Protein Labeling.
A8001 Sulfo-NHS-Biotin
K1005 EZLink Sulfo-NHS-Biotin Kit
K1151 Sulfo-NHS-Biotin Kit (90-180 kDa)
Hydrophobic Membrane-Permeable
Requires organic solvents (like DMSO) to dissolve; their membrane-permeability makes them ideal for intracellular protein tagging.
K1002 EZLink NHS-Biotin Kit
A8002 NHS-Biotin
A8010 Biotin
A8784 NHS-12-Biotin
A8013 Biotin-azide
A8007 Biotin Hydrazide
Spacer Arm Length
Standard Length
Shortest link, minimal modification size
K1002 EZLink NHS-Biotin Kit
A8002 NHS-Biotin
A8010 Biotin
A8784 NHS-12-Biotin
A8013 Biotin-azide
A8007 Biotin Hydrazide
Long Chain
The long spacer arm reduces steric hindrance, significantly improving the binding efficiency to Streptavidin beads.
A8004 NHS-LC-Biotin
A8003 Sulfo-NHS-LC-Biotin
K1003 EZLink NHS-LC-Biotin Kit
K1001 EZLink Sulfo-NHS-LC-Biotin Kit
K4401 NHS-LC-LC-Biotin Kit
A8017 NHS-LC-LC-Biotin
Functional Linkers
Cleavable Biotin
Features a disulfide bond in the spacer arm, allowing the biotin tag to be cleaved with reducing agents like DTT or BME for gentle protein elution.
A8005 Sulfo-NHS-SS-Biotin
K1004 EZLink NHS-SS-Biotin Kit
K1006 EZLink Sulfo-NHS-SS-Biotin Kit
A8006 NHS-SS-Biotin
AviTag & Magnetic Beads
Site-Specific & Reversible
Protein enrichment/purification, pull‑down for protein–protein interactions, WB/MS downstream analysis.
K4402 Biotin Labeling Kit for Avi-tag Protein (BirA)
K4409 Biotin Labeling Kit for Avi-tag Protein (BirA)
More Products >>

Validation

Streptavidin Detection & Capture Systems

After labeling, use APExBIO’s high-purity Streptavidin products for visualization and isolation:

Selection

Selected By Features Application Proudct
Streptavidin Magnetic Beads
IP and Co-IP.
High binding capacity for large proteins/antibodies.
K1301 Streptavidin Beads T3
RNA & DNA Pull-down
Ultra-low background; ideal for nucleic acid capture.
K1302 Streptavidin Beads C3
Streptavidin Conjugates
WB & ELISA
High-sensitivity enzyme for chemiluminescence.
K1229 Streptavidin -HRP
IF & Flow Cytometry
Superior photostability
K1079 Streptavidin-Cy3
K1080 Streptavidin-Cy5
K1081 Streptavidin-FITC
K4403 Streptavidin-HyperFluor™ 488
K4404 Streptavidin-HyperFluor™ 555
K4405 Streptavidin-HyperFluor™ 594
K4406 Streptavidin-HyperFluor™ 647
Biotin-tyramide
Tyramide Signal Amplification(TSA)
Detection of low-abundance targets in IHC, ICC, ISH.
A8011 Biotin Tyramide Reagent
A8012 Biotin-XX Tyramide Reagent
More Products >>

Validation

Frequently Asked Questions

Q: How do I choose between NHS-Biotin and Sulfo-NHS-Biotin?
A: Choose NHS-Biotin for organic solvent compatibility or cell-permeable applications. Choose Sulfo-NHS-Biotin for aqueous reactions and cell surface labeling where membrane permeability must be avoided.
Q: How can I elute biotinylated proteins from Streptavidin beads?
A: Use Sulfo-NHS-SS-Biotin for gentle elution via reducing agents (DTT). For non-cleavable biotin, use SDS-PAGE loading buffer and boil at 95°C, or use Desthiobiotin for competitive elution.
Q: What is the optimal Biotin-to-Protein ratio for labeling?
A: Typically, a 20-fold molar excess of biotin reagent is used to achieve a labeling ratio of 1–3 biotin molecules per protein.

Peer-Reviewed Citations

APExBIO's Biotin-Streptavidin system is validated in high-impact journals:

Bide Tong, Xiaoguang Zhang, et al. "Extracellular vesicle-based targeted protein degradation platform for multiple extracellular proteins."
EMBO Molecular Medicine. 2026 Jan 12. PMID: 41526766 (A8001)

Chao Wei, Wei Jiang, et al. "Brain endothelial GSDMD activation mediates inflammatory BBB breakdown."
Nature. 2024 May;629(8013):893-900. PMID: 38632402 (A8001)

Benson M. George, Maria Eleftheriou, et al. "Treatment of acute myeloid leukemia models by targeting a cell surface RNA - binding protein."
Nature Biotechnology (2025) (A8005)

Neuza Domingues , Steve Catarino, et al. "Connexin43 promotes exocytosis of damaged lysosomes through actin remodelling."
EMBO Journal 2024 Jul 23. PMID: 39044100 (A8005)

Fang Ye, Zhikang Yuan, et al. "Endocytic activation and exosomal secretion of matriptase stimulate the second wave of EGF signaling to promote skin and breast cancer invasion."
Cell Reports 2024 Apr 23;43(4):114002. PMID: 38547126 (A8005)

Shraddha Subramanian, Alexes C. Daquinag, et al. "Characterization of peptides targeting metastatic tumor cells as probes for cancer detection and vehicles for therapy delivery."
Cancer Research 2021 Nov 15;81(22):5756-5764. PMID: 34607842 (K1001)

Songzi Liu, Xiaoge Zhang, et al. "Mammalian IRE1α dynamically and functionally coalesces with stress granules."
Nature Cell Biology 2024 Jun;26(6):917-931. PMID: 38714852 (A8011)

Tzu-Yuan Chiu, Daniel C. Lazar, et al. "Chemoproteomic development of SLC15A4 inhibitors with anti-inflammatory activity."
Nature Chemical Biology 2024 Jan 8. PMID: 38191941 (A8011)

Wei Qin, Samuel A. Myers, et al. "Spatiotemporally-resolved mapping of RNA binding proteins via functional proximity labeling reveals a mitochondrial mRNA anchor promoting stress recovery."
Nature Communications 2021 Aug 17;12(1):4980. PMID: 34404792 (A8011)