ALFA Selector CE (Incl. 10 mg elution peptide)

Cat No: N1512 Category:

275,00 445,00 

ALFA Selector CE (for “Cold Elutable”) is an innovative affinity resin (Agarose & Magnetic) designed for the highly efficient purification of ALFA-tagged proteins in physiological buffers at low temperatures (e.g., 4°C). Each unit includes 10 mg of ALFA elution peptide.

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ALFA Selector CE (for “Cold Elutable”) is an innovative affinity resin designed for the highly efficient purification of ALFA-tagged proteins at low temperatures in physiological buffers. Please look to our Technology Section for more information about the ALFA system.

ALFA Selector CE is based on a medium-affinity (Kd ≈ 100 nM) single-domain antibody (sdAb) that is covalently immobilized on 4% cross-linked agarose or magnetic agarose beads. ALFA Selector CE is optimized for an efficient, competitive elution using physiological buffers at 4 °C using ALFA elution peptide. For this purpose, every unit of ALFA Selector CE 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, they can be purchased here.

Alternatively, elution can be performed under acidic or denaturing conditions. ALFA Selector CE can be regenerated using acid or basic conditions (100 mM Glycine pH 2.2, 150 mM NaCl, or 100 mM NaOH, respectively) and does not lose capacity even after 10 regeneration cycles. 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 CE thus features high specificity and superior capacity for ALFA fusion proteins while showing negligible non-specific background. ALFA Selector CE is compatible not only with physiological buffers but also with high stringency buffers (see ALFA Selector Manual). ALFA Selector CE thus provides great freedom to adjust the binding and washing conditions to the experimental needs.

Variations:
Conjugation Amount Cat No. RRID
4% cross-linked agarose 2000 μl N1512-L AB_3075992
4% cross-linked magnetic agarose 2000 μl N1517 AB_3075995
Related Products:

ALFA Selector ST (Super Tight), magnetic & agarose

ALFA Selector PE (Peptide Elutable), magnetic and agarose

ALFA elution peptide (10 mg)

 

Clone: C1
Host: Alpaca
Produced in: E.coli
Application: IP
Dilution: -
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)
  4. Lin C, Feng S, DeOliveira CC, Crane BR. Cryptochrome-Timeless structure reveals circadian clock timing mechanisms. Nature. 2023;617(7959):194-199. doi:10.1038/s41586-023-06009-4 (CoIP; fruit fly S2 cells)
  5. Tao X, Zhao C, MacKinnon R. Membrane protein isolation and structure determination in cell-derived membrane vesicles. Proc Natl Acad Sci U S A. 2023;120(18):e2302325120. doi:10.1073/pnas.2302325120 (vesicle purification; Sf9 cells)
Notice: To be used in vitro/ for research only. Non-toxic, non-hazardous, non-infectious.
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