ALFA Catcher PE

Catalog No. ABIN7272857

Quick Overview for ALFA Catcher PE (ABIN7272857)

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

Specificity: Recognizes the ALFA-tag (SRLEEELRRRLTE)

Characteristics: ALFA Catcher 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 Catcher PE is optimized for an efficient, competitive elution under physiological conditions using ALFA elution peptide. For this purpose, every unit of ALFA Catcher 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 Catcher 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).

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 5 to 10
• > 1 % Triton X-100, > 1% DDM, > 1 % CHAPS, > 0.5 % Deoxycholate, not compartible with SDS
• > 3 M NaCl, > 1 M MgSO4
• 3 M urea, > 2 M Guanidin-HCl
• > 100 mM DTT

Comment:

ALFA Catcher PE thus features high affinity and superior capacity for ALFA fusion proteins while showing negligible non-specific background. ALFA Catcher PE is compatible not only with physiological buffers but also with high stringency buffers. ALFA Catcher PE 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.