Biotin-Actin
α-actin from skeletal muscle, rabbit
Cat. #: 8109-01
Applications
Biotin-Actin is optimized for streptavidin/avidin-based capture and surface immobilization of actin for single-filament assays and reconstituted cytoskeletal systems. Typical uses include TIRF microscopy, tethered filament/seed assays, actin network assembly on functionalized surfaces, pull-down workflows, and biotin-dependent immunodetection formats where controlled orientation and robust anchoring are required.
Description
| Protein | Biotin-Actin |
| Origin | Skeletal muscle α-actin, rabbit |
| Molecular mass | 42 kDa |
| Protein description |
Biotin-Actin is covalently biotinylated monomeric actin (G-actin) prepared from rabbit skeletal muscle α-actin. The modification uses a long spacer arm (≈20 Å) to reduce steric hindrance and to keep the biotin moiety accessible for high-affinity avidin/streptavidin capture and biotin-dependent detection. The product is quality-controlled for polymerization competence and is validated to assemble into F-actin under standard salt/Mg2+ induction conditions (e.g., KCl/MgCl2), enabling direct use in reconstitution assays. |
| Conjugation ratio | Degree of labeling (DOL) ≈ 1 biotin per actin monomer |
Properties
| Form | Lyophilized, ready-to-use. |
| Quantity per unit | 2 × 100 µg |
| Buffer | 2 mM Tris-Cl pH 8.2, 0.4 mM ATP, 0.1 mM DTT, 0.2 mM CaCl2, 1 mM NaN3, 0.5% disaccharides (upon reconstitution with 100 µl ultrapure water to obtain 1 mg/ml) |
| Purity | >99% by scanning densitometry. |
| Purification notes | Purified by gel permeation chromatography from rabbit skeletal muscle. |
| Protein concentration | Determined by the Biuret method. |
| Storage instructions | Store at −70°C upon arrival. After reconstitution keep the protein on ice and use within 1 week. Avoid refreezing. For critical assays, clarify the working stock by gel filtration/dialysis and/or high-speed centrifugation to remove aggregates. |
| Shipping conditions | At ambient temperature. Upon delivery store at −70°C. |
| Remarks | For actin buffer exchange/dilution use MonoMix (Cat. no.: 5100-0*). In the absence of nucleators, polymerization can be initiated using PolyMix (Cat. no.: 5000-0*). For biotin-specific detection, avidin/streptavidin-based conjugates can be used under standard immunoblot conditions. For Use in Research only. Not for Use in Diagnostic Processes. |
| CAS no. |
Proteins from Hypermol® are made of the ultrapure reagents in Milli-Q™ water, as described in our publications.
Further Information
Material and Safety Data Sheet
References
Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin.
Kelley CF, Litschel T, Schumacher S, Dedden D, Schwille P, Mizuno N. eLife. 2020 Jul 13;9:e56110. doi: 10.7554/eLife.56110.
Reconstitution of contractile actomyosin rings in vesicles.
Litschel T, Kelley CF, Holz D, et al. Nat Commun. 2021 Apr 15;12(1):2254. doi: 10.1038/s41467-021-22422-7.
Product DataSheetMaterial and Safety Data Sheet
Protein Sequence on NCBI
Phosphoinositides regulate force-independent interactions between talin, vinculin, and actin.
Kelley CF, Litschel T, Schumacher S, Dedden D, Schwille P, Mizuno N.; Elife. 2020 Jul 13;9:e56110. doi: 10.7554/eLife.56110. PMID: 32657269; PMCID: PMC7384861.
Reconstitution of contractile actomyosin rings in vesicles.
Litschel, T., Kelley, C.F., Holz, D. et al.; Nat Commun 12, 2254 (2021). https://doi.org/10.1038/s41467-021-22422-7
