Anti-V5 Magnetic Beads – Catalog #B2025667

Anti-V5 Magnetic Beads (Catalog #B2025667) are affinity purification reagents that combine a magnetic core with antibodies targeting the V5 epitope tag. V5 is a 14 amino acid peptide sequence derived from the paramyxovirus SV5, widely used in molecular and cell biology as a small, immunogenic affinity tag. These beads provide one-step immunomagnetic capture of V5-fusion proteins with minimal sample handling.

Scientific Overview

The V5 epitope tag is a 14 amino acid peptide (GKPIPNPLLGLDST) recognized by widely used anti-V5 monoclonal antibodies. V5 is small enough to be appended to proteins without disrupting function, yet immunogenic enough for reliable detection and purification. It is commonly fused to the C-terminus of proteins of interest, enabling rapid identification, localization, and isolation from complex mixtures.

Magnetic bead-based purification offers practical advantages: no centrifugation is required, separation is rapid (minutes), and protein complexes remain intact under mild binding and wash conditions. The anti-V5 coating on these beads provides specific recognition of the V5 tag with minimal cross-reactivity to untagged cellular proteins.

Key applications include:

• One-step affinity purification of V5-tagged recombinant proteins
• Immunoprecipitation of V5-fusion complexes from whole-cell lysates
• Protein interaction mapping (co-IP experiments)
• Qualitative assessment of tag accessibility and protein expression

Usage & Handling Guidance

Gently equilibrate the beads (do not vortex) and add to your sample in binding buffer. Incubate at 4°C or room temperature with gentle rotation to allow antibody-epitope interaction. Apply a magnet to pellet beads, remove the supernatant, and perform 3–5 washes with ice-cold wash buffer. Elute bound protein with low pH buffer, urea solution, or SDS-PAGE loading buffer depending on downstream use.

• Binding buffer: PBS, TBS, or mild cell lysis buffer (e.g., 1% Triton X-100, 150 mM NaCl, 50 mM Tris pH 7.4).
• Washing: Include 0.1% detergent (Triton X-100 or Tween-20) and 150–500 mM NaCl to reduce non-specific binding.
• Elution: 100 mM glycine–HCl (pH 2.5), 6 M urea, or 1% SDS; pH 2.5 elution is mildest for downstream applications.

What You Get

• 500 µL of anti-V5 magnetic beads, ready to use
• Rapid, one-step affinity capture of V5-tagged proteins
• Magnetic format requiring no centrifugation
• For research use only (RUO)

Why Researchers Choose It

• V5 tag is widely recognized and immunogenic
• Magnetic beads eliminate centrifugation steps, reducing sample handling time
• Works in a broad range of lysis and binding buffers
• Compatible with downstream mass spectrometry and protein assays

Frequently Asked Questions (FAQ)

• What is the V5 epitope tag?
  The V5 epitope is a 14 amino acid sequence (GKPIPNPLLGLDST) derived from the paramyxovirus SV5 and recognized by widely used monoclonal antibodies.
• Can I use these beads at room temperature or must I use 4°C?
  Room temperature incubation is acceptable and often faster; 4°C is recommended to minimize non-specific binding and protein degradation during extended incubations.
• What is the binding capacity?
  Binding capacity depends on protein size and concentration. Start with 25–50 µL of beads per 1–10 µg of V5-tagged target and optimize based on your recovery.
• Can I perform ChIP with these beads?
  Yes. V5-tagged chromatin proteins can be cross-linked and immunoprecipitated with these beads; include a cross-linking step (e.g., formaldehyde) before lysis and pull-down.
• What is the shelf life?
  Store at –20°C; typical shelf life is 12–24 months. Avoid repeated freeze–thaw cycles; aliquot before freezing.

References

• Zheng X, Pincus D. Serial Immunoprecipitation of 3xFLAG/V5-tagged Yeast Proteins to Identify Specific Interactions with Chaperone Proteins. Bio Protoc. 2017;7(12).
• Kolodziej KE, Pourfarzad F, de Boer E, Krpic S, Grosveld F, Strouboulis J. Optimal use of tandem biotin and V5 tags in ChIP assays. BMC Mol Biol. 2009;10:6.
• Ranawakage DC, Takada T, Kamachi Y. HiBiT-qIP, HiBiT-based quantitative immunoprecipitation, facilitates the determination of antibody affinity under immunoprecipitation conditions. Sci Rep. 2019;9(1):6895.