Anti-HA Magnetic Beads – Catalog #B2025663

Anti-HA Magnetic Beads (Catalog #B2025663) are laboratory tools for rapid affinity purification of HA-tagged recombinant proteins. The beads are coated with monoclonal antibodies that bind the HA epitope (nine amino acids from influenza hemagglutinin: YPYDVPDYA), enabling one-step magnetic bead-based separation of HA fusion proteins from complex protein mixtures. Supplied as 500 μL of suspension, they offer a fast, convenient alternative to column chromatography for protein purification, immunoprecipitation, and protein-protein interaction studies.

Scientific Overview

The HA epitope (YPYDVPDYA) is a nine-amino-acid tag derived from the influenza virus hemagglutinin protein. It is one of the smallest and least disruptive epitope tags used in molecular biology, making it ideal for applications where protein size and function must be preserved. The HA tag is easily recognized by well-characterized monoclonal antibodies and has been validated in thousands of studies across cell biology, developmental biology, and biochemistry.

Anti-HA magnetic beads use high-affinity monoclonal antibodies to capture HA-tagged proteins with specificity and efficiency. The beads facilitate rapid isolation of HA fusion proteins from cell lysates, making them particularly valuable for co-immunoprecipitation (co-IP) experiments, protein complex purification, and studies of transient or weak protein-protein interactions. Their small size and mild handling make them suitable for maintaining protein complex integrity during purification.

Key applications include:

• One-step affinity purification of HA-tagged proteins from lysates
• Co-immunoprecipitation of protein-protein interaction partners
• Isolation of protein complexes while preserving native interactions
• Pull-down assays in structural and cell biology
• Rapid sample preparation for mass spectrometry

Usage & Handling Guidance

These beads are supplied as a suspension. Gently mix before each use; avoid vortexing, which may damage the antibody coating. Add the beads directly to your lysate or protein mixture and incubate at 4°C for 30–60 minutes to allow binding, then place the tube in a magnetic rack for 1–2 minutes to pellet the beads. Remove the supernatant and wash the beads 3–5 times with binding buffer or PBS.

• Binding buffer: PBS, TBS, or your native lysis buffer works well. Standard lysis buffers (with protease inhibitors if desired) are compatible.
• Incubation: 30–60 minutes at 4°C is typical. Longer incubations improve recovery but may also increase co-precipitation of non-specific proteins.
• Washing: Perform 3–5 washes with 10–15 bead volumes of binding buffer per wash to remove non-specifically bound proteins.
• Elution: Elute with low pH buffer (0.1 M glycine, pH 2.8–3.0) or denaturing conditions (6 M urea, 8 M guanidinium chloride). Some applications tolerate gentler elution with hot 2% SDS.
• Magnetic separation: Use a commercial magnetic rack suitable for 1.5 or 2 mL tubes.

What You Get

• 500 μL of anti-HA magnetic beads in suspension
• Ready-to-use beads for immediate co-immunoprecipitation and purification
• High-specificity monoclonal antibodies against the HA epitope
• For research use only (RUO)

Why Researchers Choose It

• HA is a small, well-tolerated epitope tag with minimal impact on protein folding and function
• Magnetic bead format enables rapid purification without column chromatography
• Monoclonal antibodies provide excellent specificity and low background
• Works well for co-immunoprecipitation and protein complex isolation
• Compatible with downstream applications including mass spectrometry

Frequently Asked Questions (FAQ)

• How specific is the anti-HA antibody?
  Monoclonal anti-HA antibodies are highly specific to the HA epitope (YPYDVPDYA) and show negligible cross-reactivity with other epitope tags or endogenous proteins in cell lysates.
• Can I use these beads with live cells or only with lysates?
  These beads are designed for use with cell lysates and purified protein. For whole-cell immunofluorescence, use anti-HA antibodies without magnetic beads.
• What is the recovery efficiency?
  Recovery depends on incubation time, antibody-epitope affinity, and bead concentration. Typical recoveries are 50–90% under optimized conditions. Request a COA/TDS for lot-specific performance data.
• Can I scale up the protocol?
  Yes. These beads scale linearly; simply use proportionally more beads and longer incubation times. For large-scale purifications (>10 mL lysate), consider contacting us for bulk quantities and optimization advice.
• How do I remove the beads from my sample?
  Place the tube in a magnetic rack for 1–2 minutes and carefully pipette off the supernatant. Beads will adhere to the tube wall. A second brief incubation in the magnet can help ensure complete removal.

References

• 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;10(1):4403.
• Liu D, Liu C, Hu J, Hang L, Li X, Wei Y, Zhu H, Zhang Q, Wang X. Construction and evaluation of HA-epitope-tag introduction onto the VP1 structural protein of a novel HY12 enterovirus. Virology. 2018;525:106-116.
• Zhao X, Li G, Liang S. Several affinity tags commonly used in chromatographic purification. J Anal Methods Chem. 2013;2013:581093.