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Immunostaining of HeLa cells expressing an ALFA-Vimentin fusion protein with an anti-ALFA sdAb (Cat. No. N1505-DBCO, dilution 1:500) bearing a 10-nucleotide single-stranded DNA, imaged by 3D DNA-PAINT. The histogram refers to the region marked by a small yellow rectangle where 2 individual vimentin filaments are resolved although being only ~90 nm apart. The localization precision was 5.2 nm. Figure modified from Götzke et al. Nat Commun 2019.

sdAb anti-ALFA

Cat No: N1505 Category:

Price range: 275,00 € through 625,00 €

In-house developed single-domain antibody (sdAb) that recognizes the small rationally-designed ALFA-tag (SRLEEELRRRLTE) with low pM affinity.

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The ALFA-tag is a novel, rationally designed epitope tag (SRLEEELRRRLTE) and is part of NanoTag’s proprietary technology. In contrast to most conventional epitope tags, the ALFA-tag was developed based on rational considerations, which enabled us to equip it with desired features (learn more about the ALFA system here). The ALFA-tag, in combination with three sdAbs, provides a unique platform for employing a single epitope tag in virtually all biochemical and biomedical applications. A complete characterization of the system can be found in an open-source article published in Nat. Commun. in 2019.

This sdAb (N1505) is the clone 1G5, which is the member of the ALFA system family with the strongest affinity towards ALFA-tag (~26 pM). It is the exact clone also used in our FluoTag®-X2 anti-ALFA (N1502), recombinant anti-ALFA antibody (rHcAb; N1582-N1586), anti-ALFA bispecifics (N1590-N1592), sdAb anti-ALFA Halo fusion (N1541), and the same one as in the ALFA Selector ST (N1511).

MSDS
sdAb unconjugated
sdAb-Biotin/DBCO
sdAb-HRP
Protocols
Reconstitution and Storage
Variations:
Conjugation Amount Cat No. RRID
unconjugated C-terminal Cys 250 μg N1505-250ug AB_3075986
Biotin 250 μg N1505-Biotin AB_3075987
DBCO 250 μg N1505-DBCO AB_3075988
HRP 200 μL N1505-HRP AB_3075989
Related Products:

FluoTag®-X2 anti-ALFA (Cat. No. N1502)

sdAb anti-ALFA Halo fusion (Cat. No. 1541)

Recombinant anti-ALFA antibody (rHcAb) (Cat. No. N1582 – N1586)

anti-ALFA Bispecifics (Cat. No. N1590 – N1592)

View related products
Clone: 1G5
Host: Alpaca
Produced in: E.coli
Application: IF, WB
Dilution: ELISA with sdAb anti-ALFA HRP 1:5000
Capacity: N/A
Antigen: -
Targets: ALFA-tag
Specificity: Recognizes the ALFA-tag (SRLEEELRRRLTE).
Formulation:

DBCO- and Biotin-conjugated sdAbs (1:1 labeling on the C-terminus) are lyophilized from PBS pH 7.4. Unconjugated sdAbs are lyophilized from 5 mM KPi pH 6.0, 300 mM NaCl, 0.5 mM EDTA pH 6.0. Reconstitute with 250 µL pure deionized water for immediate use or with 250 µL sterile 50% glycerol for storage at -80°C.

The HRP-conjugated sdAbs are lyophilized from HRP protector buffer and should be reconstituted with 200 µL pure deionized water for storage at 4°C or with 200 µL high purity, sterile 50% glycerol for storage at -80°C.

For more details, click the Protocols button above and check Reconstitution and Storage.
kDa: 16.0
Ext Coef: 18638
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. 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. Honsa M, Pachmayr I, Heinze L, et al. Imaging Ligand-Receptor Interactions at Single-Protein Resolution with DNA-PAINT. Small Methods. Published online April 3, 2025. doi:10.1002/smtd.202401799 (IF; DNA-PAINT; A549 cells)
  3. Cheetham-Wilkinson IJ, Sivalingam B, Flitton C, et al. RpH-ILV: Probe for lysosomal pH and acute LLOMe-induced membrane permeabilization in cell lines and Drosophila. Sci Adv. 2025;11(1):eadr7325. doi:10.1126/sciadv.adr7325 (N1505-Biotin; WB; HEK293T; fruit fly S2 cells)
  4. Kalogriopoulos NA, Tei R, Yan Y, et al. Synthetic GPCRs for programmable sensing and control of cell behaviour. Nature. Published online December 4, 2024. doi:10.1038/s41586-024-08282-3 (N1505-Biotin; magnetic-activated cell sorting MACS; primary human T cells)
  5. Mougios N, Cotroneo ER, Imse N, et al. NanoPlex: a universal strategy for fluorescence microscopy multiplexing using nanobodies with erasable signals. Nat Commun. 2024;15(1):8771. Published 2024 Oct 10. doi:10.1038/s41467-024-53030-w
  6. Shaib AH, Chouaib AA, Chowdhury R, et al. One-step nanoscale expansion microscopy reveals individual protein shapes. Nat Biotechnol. Published online October 9, 2024. doi:10.1038/s41587-024-02431-9
  7. Hellmeier J, Strauss S, Xu S, et al. Quantification of absolute labeling efficiency at the single-protein level. Nat Methods. Published online April 24, 2024. doi:10.1038/s41592-024-02242-5 (ICC/IF; DNA-PAINT)
  8. Dodd DO, Mechaussier S, Yeyati PL, et al. Ciliopathy patient variants reveal organelle-specific functions for TUBB4B in axonemal microtubules Science. 2024;384(6694):eadf5489. doi:10.1126/science.adf5489 (WB; IMCD3 cells)
  9. van Zwam MC, Dhar A, Bosman W, et al. IntAct: A nondisruptive internal tagging strategy to study the organization and function of actin isoforms. PLoS Biol. 2024;22(3):e3002551. Published 2024 Mar 11. doi:10.1371/journal.pbio.3002551 (N1505-HRP; WB; HT1080 cells)
  10. 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 (WB; C. elegans)
  11. Igreja C, Loschko T, Schäfer A, et al. Application of ALFA-Tagging in the Nematode Model Organisms Caenorhabditis elegans and Pristionchus pacificus. Cells. 2022;11(23):3875. Published 2022 Dec 1. doi:10.3390/cells11233875 (WB; C. elegans, P. pacificus)
  12. 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 (WB; yeast)
  13. Buglino JA, Ozakman Y, Xu Y, Chowdhury F, Tan DS, Glickman MS. Diisonitrile Lipopeptides Mediate Resistance to Copper Starvation in Pathogenic Mycobacteria. mBio. 2022;13(5):e0251322. doi:10.1128/mbio.02513-22 (WB; M. tuberculosis)
  14. 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 (WB; HeLa infected with Y. enterocolitica)
  15. 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 (Homodimer formation)
  16. 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 (WB; fruit fly)
  17. Zhang Q, Miyamoto A, Watanabe S, et al. Engineered fast-dissociating antibody fragments for multiplexed super-resolution microscopy. Cell Rep Methods. 2022;2(10):100301. Published 2022 Sep 20. doi:10.1016/j.crmeth.2022.100301 (IF, multiplexed super-resolution microscopy; Xenopus laevis XTC cells)
  18. 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 (WB; yeast)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
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