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sdAb anti-Halo clone 1A12_kinetic analysis_NanoTag Biotechnologies

Kinetic analysis of the interaction between immobilized sdAb anti-Halo clone 1A12 (biotinylated, 5 µg/mL) and a Halo-tagged analyte (20–0.3125 nM), measured by Bio-Layer Interferometry (BLI) using Streptavidin biosensors. Association and dissociation phases (colored traces) were globally fitted (black lines) using a 1:1 binding model. The resulting kinetic parameters indicate high-affinity binding with an association rate constant kon = (2.96 ± 0.17) × 10⁵ M⁻¹ s⁻¹ and a dissociation rate constant koff = (5.21 ± 0.01) × 10⁻⁵ s⁻¹ corresponding to an equilibrium dissociation constant KD = 176.9 ± 0.2 pM.

sdAb anti-Halo

Cat No: N5005 Category:

Price range: 290,00 € through 655,00 €

sdAb anti-Halo is an in-house developed single-domain antibody (sdAb) that specifically binds HaloTag®. It enables highly specific detection of Halo fusion proteins and is generally compatible with simultaneous Halo ligand binding.

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The HaloTag®(1) technology is a widely used and highly versatile platform in molecular and cellular biology, enabling detailed analyses of protein function, dynamics, and molecular interactions. It is based on a genetically engineered haloalkane dehalogenase derived from bacteria, which has been modified to form a specific and stable covalent bond with synthetic Halo ligands. These ligands can be functionalized with a broad array of moieties, including fluorophores, biotin, and affinity tags, enabling diverse applications in live-cell imaging, protein purification, and biochemical assays.

Unlike conventional Halo ligands, sdAb anti-Halo (clone 1A12) recognizes surface-exposed epitope on the HaloTag® and is generally compatible with simultaneous Halo ligand binding, thus preserving the functional integrity of the HaloTag® system during multiplexed applications.

We provide this product conjugated to various labels and offer custom conjugation services with most commercially available fluorophores, provided these are available in a thiol-reactive activated form.

(1) HaloTag® is a registered trademark of Promega Corporation.

MSDS
sdAb unconjugated
sdAb-Biotin/DBCO
Protocols
Reconstitution and Storage
Maleimide Labeling
IF (ICC) Protocol
Variations:
Conjugation Amount Cat No. RRID
unconjugated C-terminal Cys 250 μg N5005-250ug AB_3714728
Biotin 250 μg N5005-Biotin AB_3714729
DBCO 250 μg N5005-DBCO AB_3714730
Related Products: View related products
Clone: 1A12
Host: Alpaca
Produced in: E.coli
Application: ELISA, IF
Dilution: N/A
Capacity: N/A
Antigen: HaloTag
Targets: HaloTag
Specificity: Recognizes the HaloTag. No known cross-reactivity to other proteins.
Formulation:

DBCO and biotin conjugated sdAbs (1:1 labeling on the C-term.) are lyophilized from PBS pH 7.4. Unconjugated sdAbs are lyophilized from 10 mM KPi,150 mM NaCl, 1 mM EDTA pH 6.0. Reconsitute with 250 µL pure water for immediate use or with 250 µL 50% glycerol for storage at -80 °C. For more details, click the Protocols button above and check Reconstitution and Storage.
kDa: 14.9
Ext Coef: 20128
Shipping: Ambient temperature
Storage:

Vials containing lyophilized reagent can be stored at 2-8°C for up to 12 months. After reconstitution, store at -80°C for up to 6 months. Working aliquots can be stored at -20°C for up to 4 weeks. For more details, click the Protocols button above and check Reconstitution and Storage.
Protocols:

Relevant protocols can be found under the Protocols button above. For additional information, visit our Resources page.
References:
  1. Los GV, Encell LP, McDougall MG, et al. HaloTag: a novel protein labeling technology for cell imaging and protein analysis. ACS Chem Biol. 2008;3(6):373-382. doi:10.1021/cb800025k
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
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