Product Description
EnkiBeadsTM Protein A (or A/G) Antibody Purification Kit utilizes EnkiLife's proprietary protein coupling technology to covalently immobilize Protein A (or A/G) onto NHS-activated superparamagnetic microspheres. Compared to similar products on the international market, this kit offers higher antibody binding capacity and lower non-specific protein adsorption, with more uniform elution conditions. A single step purification yields >90% pure antibody from serum samples.
This product features nanoscale magnetic microspheres with an ultra-large specific surface area, significantly reducing antibody binding time. Skilled users can complete the antibody binding process within 15 minutes and the entire purification workflow within 30 minutes. The kit includes pre-optimized buffer formulations that provide optimal reaction conditions for antibody purification, ensuring enhanced protocol stability and reproducibility.
The product is reusable and suitable for antibody purification from plasma, ascites, tissue culture supernatant, and other sample types. It can also be used for antibody immobilization and related research applications. Users should select the appropriate bead type (Protein A or Protein A/G) based on the species and isotype of the target antibody. Refer to Appendix 1 for the affinity comparison between Protein A/G beads and various antibodies.
Storage Conditions and Shelf Life
Storage Conditions: Store at 2–8°C. DO NOT FREEZE.
Shelf Life: 2 years
Precautions
Requires a magnetic separator (see Appendix 3 for compatible models).
Resuspend beads completely before each use by vortexing.
Store beads in supplied buffers—do not allow to dry.
DO NOT freeze or centrifuge beads—this causes irreversible aggregation.
Research Use Only (RUO)—not for diagnostic or therapeutic use.
Frequently Asked Questions (FAQ)
Q1: How can I improve antibody binding efficiency?
A1: Binding efficiency depends on antibody species/isotype and Protein A/G affinity (see Appendix 1). For low-affinity antibodies:
1. Increase incubation time (30–120 min)
2. Raise binding buffer pH (8–9)
3. Reduce ionic strength (25–100 mM NaCl)
4. Switch to Protein A/G beads for broader affinity.
Q2: How can I improve antibody elution efficiency?
A2: High-affinity interactions may reduce elution. Try:
Lower elution buffer pH (1.9–2.5)
Increase ionic strength (use 2–3 M MgCl₂)
Extend elution time.
Note: Low pH may cause aggregation—neutralize immediately with Tris or HEPES.
Q3: How do I prevent bead aggregation during storage/use?
A3: Store at 2–8°C. Add 0.1% (v/v) non-ionic detergent (NP-40, Tween-20, or Triton X-100) to Binding and Elution Buffers. If aggregation occurs after low-pH elution, neutralize with Binding Buffer and resuspend in Tris buffer (pH 7.5) containing 0.1% Tween-20, then sonicate for 2 minutes in a water bath. This does not affect binding capacity.
Q4: Beads stick to tube walls—how to prevent?
A4: Use low-protein-binding tubes. Add 0.01–0.1% (v/v) non-ionic detergent to buffers to reduce adhesion.
Q5: Beads clump together during use?
A5: Clumping occurs when beads remain on magnet too long. Use brief sonication (2 min) to disperse. WARNING: Do NOT sonicate after sample addition and before elution—this will release bound antibodies.
