ALFA Selector ST

Cat No: N1511 Category:

275,00 445,00 

ALFA Selector ST (for “Super Tight”) is an innovative, high-affinity resin (Agarose & Magnetic) designed for efficient and clean pull-down of ALFA-tagged proteins.

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ALFA Selector ST (for “Super Tight”) is an innovative, high-affinity resin designed for the highly efficient pull-down of ALFA-tagged proteins.

Please look to our Technology Section for more information about the ALFA system.

ALFA Selector ST is based on a single-domain antibody (sdAb), which is covalently immobilized on 4% cross-linked agarose or agarose-magnetic beads and binds ALFA-tagged proteins with extraordinarily high affinity (Kd = 26 pM). Target proteins are efficiently released from ALFA Selector ST under acidic (e.g., 0.1 M glycin pH2.2) or denaturing conditions. The extremely tight interaction with ALFA-tagged proteins makes competitive elution using ALFA elution peptide inefficient. We recommend using ALFA Selector PE for competitive elution under physiological and room temperature condition conditions. For efficient elutions at 4°C, we recommend using the ALFA Selector CE

The innovative, oriented and highly selective attachment of the active sdAbs via a flexible linker guarantees optimal accessibility for target proteins and at the same time largely eliminates batch-to-batch variations. Due to the single-chain nature of sdAbs and their covalent attachment, no “leakage” of light and heavy chains from IgGs is observed even during elution with strongly denaturing and reducing conditions (e.g., SDS sample buffer). ALFA Selector ST thus features high affinity and superior capacity for ALFA fusion proteins while showing negligible non-specific background.

ALFA Selector ST is compatible not only with physiological buffers but also with high stringency buffers (see ALFA Selector Manual). ALFA Selector ST thus provides great freedom to adjust the binding and washing conditions to the experimental needs.

MSDS
Variations:
Conjugation Amount Cat No. RRID
4% cross-linked magnetic agarose 2000 μl N1516-L AB_3075994
4% cross-linked agarose 2000 μl N1511-L AB_3075991
Related Products:

ALFA Selector PE (Peptide Elutable), magnetic and agarose

ALFA Selector CE (Cold Elutable), magnetic and agarose

ALFA elution peptide (10 mg)

 

Clone: 1G5
Host: Alpaca
Produced in: E.coli
Application: IP
Dilution: N/A
Capacity: > 3 µg ALFA-tagged GFP per µL of packed beads
Antigen: -
Targets: ALFA-tag
Specificity: Recognizes the ALFA-tag (SRLEEELRRRLTE)
Formulation: Resin is 50 % slurry in PBS containing 20 % Ethanol.
kDa: -
Ext Coef: -
Shipping: Ambient temperature
Storing: Store at 4 °C for up to 12 months. Do not freeze!
Protocols:

Detail protocols for IP and elutions using the ALFA system can be found here in the ALFA Selector Manual 

Protocols can be found on our Resources page.

References:
  1. Götzke H, Kilisch M, Martínez-Carranza M, Sograte-Idrissi S, Rajavel A, Schlichthaerle T, Engels N, Jungmann R, Stenmark P, Opazo F, Frey S. The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications. Nat Commun. 2019 Sep 27;10(1):4403. doi: 10.1038/s41467-019-12301-7. PMID: 31562305; PMCID: PMC6764986.
  2. Kilisch M, Götzke H, Gere-Becker M, Crauel A, Opazo F, Frey S. Discovery and Characterization of an ALFA-Tag-Specific Affinity Resin Optimized for Protein Purification at Low Temperatures in Physiological Buffer. Biomolecules. 2021;11(2):269. Published 2021 Feb 12. doi:10.3390/biom11020269 (IP; HeLa)
  3. Kohler V, Kohler A, Berglund LL, et al. Nuclear Hsp104 safeguards the dormant translation machinery during quiescence. Nat Commun. 2024;15(1):315. Published 2024 Jan 5. doi:10.1038/s41467-023-44538-8 (UV-activated site-specific crosslinking; yeast)
  4. Zhang L, Stauffer WT, Wang JS, et al. Recruitment of Polo-like kinase couples synapsis to meiotic progression via inactivation of CHK-2. Elife. 2023;12:e84492. Published 2023 Jan 26. doi:10.7554/eLife.84492 (IP; C. elegans)
  5. Mergiya TF, Gundersen JET, Kanhema T, Brighter G, Ishizuka Y, Bramham CR. Detection of Arc/Arg3.1 oligomers in rat brain: constitutive and synaptic activity-evoked dimer expression in vivo. Front Mol Neurosci. 2023;16:1142361. Published 2023 Jun 9. doi:10.3389/fnmol.2023.1142361 (IP; rat dentate gyrus)
  6. Carsten A, Rudolph M, Weihs T, et al. MINFLUX imaging of a bacterial molecular machine at nanometer resolution. Methods Appl Fluoresc. 2022;11(1):10.1088/2050-6120/aca880. Published 2022 Dec 13. doi:10.1088/2050-6120/aca880 (IP; HeLa infected with Y. enterocolitica)
  7. Ishizuka Y, Mergiya TF, Baldinotti R, et al. Development and Validation of Arc Nanobodies: New Tools for Probing Arc Dynamics and Function [published correction appears in Neurochem Res. 2022 Mar 31;:]. Neurochem Res. 2022;47(9):2656-2666. doi:10.1007/s11064-022-03573-5 (IP; human SH-SY5Y, rat dentate gyrus)
  8. Shvarev D, Schoppe J, König C, et al. Structure of the HOPS tethering complex, a lysosomal membrane fusion machinery. Elife. 2022;11:e80901. Published 2022 Sep 13. doi:10.7554/eLife.80901 (Pulldown for MS; yeast)
  9. Akhuli D, Dhar A, Viji AS, Bhojappa B, Palani S. ALIBY: ALFA Nanobody-Based Toolkit for Imaging and Biochemistry in Yeast. mSphere. 2022;7(5):e0033322. doi:10.1128/msphere.00333-22 (IP; yeast)
  10. Bloemeke N, Meighen-Berger K, Hitzenberger M, et al. Intramembrane client recognition potentiates the chaperone functions of calnexin. EMBO J. 2022;41(24):e110959. doi:10.15252/embj.2022110959 (IP; HEK293T)
  11. Kusakari K, Machida T, Ishida Y, et al. The complex formation of MASP-3 with pattern recognition molecules of the lectin complement pathway retains MASP-3 in the circulation. Front Immunol. 2022;13:907023. Published 2022 Aug 16. doi:10.3389/fimmu.2022.907023 (Protein purification; CHO)
  12. Xu J, Kim AR, Cheloha RW, et al. Protein visualization and manipulation in Drosophila through the use of epitope tags recognized by nanobodies. Elife. 2022;11:e74326. Published 2022 Jan 25. doi:10.7554/eLife.74326 (IP)
  13. Gomkale R, Linden A, Neumann P, et al. Mapping protein interactions in the active TOM-TIM23 supercomplex. Nat Commun. 2021;12(1):5715. Published 2021 Sep 29. doi:10.1038/s41467-021-26016-1 (IP; yeast)
  14. Cui Y, Ma L, Schacke S, et al. Merlin cooperates with neurofibromin and Spred1 to suppress the Ras-Erk pathway. Hum Mol Genet. 2021;29(23):3793-3806. doi:10.1093/hmg/ddaa263 (IP; HEK293)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
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