ALFA Catcher CE

Catalog No. ABIN7272856

Quick Overview for ALFA Catcher CE (ABIN7272856)

Target: ALFA-Tag

Application: Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP), Protein Complex Immunoprecipitation (Co-IP)

Bead Ligand: Antibody

Bead Matrix: Agarose beads

Bead Size: 90 μm

Product Details

Purpose: ALFA Catcher CE (for “Cold Elutable”) is an innovative affinity resin (Agarose) 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.

Specificity: Recognizes the ALFA-tag (SRLEEELRRRLTE)

Characteristics: ALFA Catcher 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 Catcher 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 Catcher 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 Catcher 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).

Components: 4 % cross-linked agarose (bead size 50-150 μm) with covalently immobilized single-domain antibody, lyophilized ALFA elution peptide

Material not included: wash buffers, columns, tubes

Application Details

Application Notes: Coating: sdAb anti-ALFA tag clone 1G5 variant
Matrix: 4 % cross-linked agarose, bead size 50-150 μm
Capacity: > 3 µg ALFA-tagged GFP per µL of packed beads (= 2 μL of slurry)
Buffer Compatibility:

• Common buffer substances at pH 6 to 10
• 0.8 % Triton X-100, 0.8 % DDM, 0.8 % CHAPS, 0.1 % Deoxycholate, not compartible with SDS
• > 3 M NaCl, 0.8 M MgSO4
• 3 M urea, 0.3 M Guanidin-HCl
• > 100 mM DTT

Comment:

ALFA Catcher CE thus features high specificity and superior capacity for ALFA fusion proteins while showing negligible non-specific background. ALFA Catcher CE is compatible not only with physiological buffers but also with high stringency buffers. ALFA Catcher CE thus provides great freedom to adjust the binding and washing conditions to the experimental needs.

Restrictions: For Research Use only

Handling

Buffer: Resin is 50 % slurry in PBS containing 20 % Ethanol. Separately, 10 mg of lyophilized ALFA elution peptide is also provided

Storage: 4 °C

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

Expiry Date: 12 months

Target Details

Target: ALFA-Tag

Target Type: Tag

Background: The ALFA-tag is a novel, highly versatile epitope tag (core sequence SRLEEELRRRLTE) that can be employed for a wide range of life science applications.