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ALFA Selector PE (Incl. 10 mg elution peptide)

Cat No: N1510 Category:

275,00 € – 445,00 €

ALFA Selector PE (for “Peptide Elutable”) is an innovative affinity resin (Agarose & Magnetic) designed for the highly efficient purification of native ALFA-tagged target proteins under physiological conditions. Each unit includes 10 mg of ALFA elution peptide.

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ALFA Selector PE (for “Peptide Elutable”) is an innovative affinity resin designed for the highly efficient purification of native ALFA-tagged target proteins under physiological conditions. Please look to our Technology Section for more information about the ALFA system.

ALFA Selector PE is based on a high-affinity (Kd = 11 nM) single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose or magnetic agarose beads. ALFA Selector PE is optimized for an efficient, competitive elution under physiological conditions using ALFA elution peptide. For this purpose, every unit of ALFA Selector PE is supplied with 10 mg of ALFA elution peptide, which is sufficient for 25 mL of elution buffer. If more ALFA elution peptide is needed, it can be purchased here.

Efficient elution using ALFA elution peptide should be performed at room temperature, if you need to elute in the cold (e.g., 4°C), Please use ALFA Selector CE (for “Cold Elutable”).

Alternatively, elution can be performed under acidic or denaturing conditions. The innovative, oriented and selective attachment via a flexible linker guarantees high accessibility of the sdAbs and 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 during elution, even under harsh conditions (e.g., SDS sample buffer).

ALFA Selector PE thus features high affinity and superior capacity for ALFA fusion proteins while showing negligible non-specific background. ALFA Selector PE is compatible not only with physiological buffers but also with high stringency buffers (see ALFA Selector Manual). ALFA Selector PE thus provides great freedom to adjust the binding and washing conditions to the experimental needs.

MSDS
Protocols
ALFA Selector Manual
ALFA Elution Peptide
Variations:
Conjugation Amount Cat No. RRID
4% cross-linked agarose 2000 μl N1510-L AB_3075990
4% cross-linked magnetic agarose 2000 μl N1515-L AB_3075993
Related Products:

ALFA Selector ST (Super Tight), magnetic & agarose

ALFA Selector CE (Cold Elutable), magnetic and agarose

ALFA elution peptide (10 mg)

 
Clone: 1G5 variant
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. Separately, 10 mg of lyophilized ALFA elution peptide is also provided.
kDa: -
Ext Coef: -
Shipping: Ambient temperature
Storing:

Store at 4 °C for up to 12 months. Do not freeze!
Protocols:

Please dissolve the lyophilized peptide (10 mg) in 250 µl of distilled water. This results in an ALFA peptide concentration of 40 mg/ml or 20 mM (100x).

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 
  3. Krauter D, Stausberg D, Hartmann TJ, et al. Targeting PI3K/Akt/mTOR signaling in rodent models of PMP22 gene-dosage diseases. EMBO Mol Med. Published online February 21, 2024. doi:10.1038/s44321-023-00019-5 (IP; rat sciatic nerve, HEK293T)
  4. Sanfilippo P, Kim AJ, Bhukel A, et al. Mapping of multiple neurotransmitter receptor subtypes and distinct protein complexes to the connectome. Neuron. Published online January 13, 2024. doi:10.1016/j.neuron.2023.12.014 (IP, MS; fruit fly)
  5. 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 (IP; P. pacificus)
  6. Zhao J, Makhija S, Zhou C, et al. Structural insights into the human PA28-20S proteasome enabled by efficient tagging and purification of endogenous proteins. Proc Natl Acad Sci U S A. 2022;119(33):e2207200119. doi:10.1073/pnas.2207200119 (Protein purification; human Expi293F)
  7. Jauslin T, Lamrabet O, Crespo-Yañez X, et al. How Phagocytic Cells Kill Different Bacteria: a Quantitative Analysis Using Dictyostelium discoideum. mBio. 2021;12(1):e03169-20. Published 2021 Feb 16. doi:10.1128/mBio.03169-20 (IP; D. discoideum)
  8. Siwach M, Kumar L, Palani S, et al. An organelle-tethering mechanism couples flagellation to cell division in bacteria. Dev Cell. 2021;56(5):657-670.e4. doi:10.1016/j.devcel.2021.01.013 (IP; Caulobacter)
  9. Guilhen C, Lima WC, Ifrid E, Crespo-Yañez X, Lamrabet O, Cosson P. A New Family of Bacteriolytic Proteins in Dictyostelium discoideum. Front Cell Infect Microbiol. 2021;10:617310. Published 2021 Feb 3. doi:10.3389/fcimb.2020.617310 (IP; D. discoideum)
  10. Perego E, Reshetniak S, Lorenz C, et al. A minimalist model to measure interactions between proteins and synaptic vesicles. Sci Rep. 2020;10(1):21086. Published 2020 Dec 3. doi:10.1038/s41598-020-77887-1 (Protein purification, human HEK293)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
Legal terms: By purchasing this product you agree to our general terms and conditions.

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