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| Product Name | Anti-HA Magnetic Beads |
| Catalog No. | MAGBEA-0013 |
| Description | Anti-HA monoclonal antibody-conjugated superparamagnetic beads for immunoprecipitation and purification of HA epitope-tagged recombinant proteins. The anti-HA antibody, recognizing the YPYDVPDYA peptide derived from influenza hemagglutinin, is covalently immobilized through oriented coupling chemistry. Competitive elution with HA peptide under native conditions preserves protein complexes and enzymatic activity. |
| Intended Use | Immunoprecipitation and small-scale purification of HA-tagged (YPYDVPDYA) recombinant proteins from mammalian, insect, yeast, and bacterial expression systems for protein interaction analysis and functional characterization. |
| Principle / Technology | Monoclonal anti-HA antibody specific for the YPYDVPDYA epitope is covalently conjugated to the bead surface via Fc-region-oriented coupling. The antibody recognizes the HA tag at N-terminal, C-terminal, or internal positions with sequence specificity. After target capture and washing, HA-tagged proteins are competitively eluted with free HA peptide under physiological pH conditions. |
| Detection Method | BCA and UV280 for antibody density; HA-GFP binding isotherm; SDS-PAGE and anti-HA Western blot; mass spectrometry; enzymatic activity assay of eluted proteins |
| Sample Type | HA-tagged proteins from E. coli, S. cerevisiae, P. pastoris, baculovirus/insect cells, and mammalian cells (HEK293, CHO, HeLa); compatible with N-terminal, C-terminal, and internal HA fusions; whole cell lysates and nuclear/cytoplasmic fractions |
| Performance Range / Specifications | Bead diameter: 1-2 µm; antibody density: 2-4 µg/mg; binding capacity: 0.3-1.5 mg HA-GFP/mL settled beads; Kd ~10^-9 M; elution >85% with HA peptide; non-specific binding <3%; magnetic separation <10 seconds |
| Sensitivity / LOD | IP from lysates with <10 ng/mL HA-tagged target; single-step enrichment sufficient for MS analysis; near-endogenous level detection of HA-tagged factors; linear binding across 0.1-100 µg input range |
| Specificity | Specific YPYDVPDYA recognition; no cross-reactivity with FLAG, myc, His6, V5, T7, GST, or MBP tags; minimal endogenous protein binding confirmed by negative control; position-independent tag recognition |
| Reaction Conditions / Protocol | Equilibrate beads in TBS; add pre-cleared lysate; incubate 1-2 hours at 4°C with rotation; magnetic separation; wash 3-5 times with lysis buffer; wash with TBS; elute with 0.5-1 mg/mL HA peptide in TBS for 15-30 minutes; separate beads; collect eluate; total protocol: 2-3 hours |
| Components / Formulation | Anti-HA monoclonal antibody-conjugated superparamagnetic beads, 25-50% slurry in PBS pH 7.4 with 0.02% sodium azide; HA peptide available separately |
| Storage Conditions | 2-8°C in PBS/azide; do not freeze; protect from light; vortex gently before use; use aseptic technique |
| Shelf Life | 12 months at 2-8°C; antibody activity >85% at expiry; preservative effective; opened: 6 months |
| Package Specifications | 0.2 mL, 0.5 mL, 1 mL settled bead volume; sufficient for 5-50 IP reactions; custom packaging available |
| Product Form | Dark brown bead suspension; homogeneous after vortexing; clear supernatant after magnetic separation; no visible aggregates |
| Quality Control | Per lot: antibody density, HA-GFP binding capacity, elution efficiency, non-specific binding, antibody leaching, endotoxin (<0.1 EU/mg), sterility, functional IP validation |
| Key Features | Competitive HA peptide elution preserves complexes; oriented antibody coupling; low non-specific binding; validated for co-IP and MS applications; clone HA.11 equivalent specificity |
| Purity | Antibody >95% by SDS-PAGE; no protease activity; endotoxin <0.1 EU/mg; sodium azide verified |
| Concentration | 25-50% slurry; antibody 2-4 µg/mg; binding capacity 0.3-1.5 mg/mL settled beads |
| Activity / Unit Definition | HA-GFP binding: 0.3-1.5 mg/mL; Kd ~10^-9 M; elution >85%; regenerable 2-3 times |
| Molecular Weight | Anti-HA IgG: ~150 kDa; HA peptide: ~1,102 Da |
| Source / Origin | Mouse monoclonal anti-HA antibody; synthetic polymer-magnetite beads; antibody purified by Protein A/G; animal-derived component traceable |
| pH Range / Optimal pH | Binding: pH 6.5-8.5 (optimal 7.0-7.5); competitive elution at physiological pH; antibody stable pH 5-9; bead integrity pH 3-11 |
| Shipping Conditions | Cold pack at 2-8°C; do not freeze; non-hazardous; standard courier |
| Expiration Date / Stability | 12 months at 2-8°C; real-time stability monitored; binding >85% at expiry; no antibody leaching increase |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic use; mouse monoclonal; ISO 9001 manufacturing |
| Compatibility | Compatible with RIPA, NP-40, Triton X-100; 150-500 mM NaCl; DTT up to 2 mM; EDTA up to 5 mM; SDS <0.05%; HA peptide compatible with downstream enzymatic assays |
| Recommended Buffer System | Storage: PBS pH 7.4, 0.02% NaN3; binding/wash: TBS, 0.1% NP-40; elution: TBS + 0.5-1 mg/mL HA peptide; neutral pH elution preserves protein integrity |
| Application Notes / Precautions | Pre-clear lysate; optimize bead-to-protein ratio; extend elution to 1 hour for high-affinity binders; HA peptide can be reused 2-3 times; remove peptide by dialysis if needed for downstream applications |
| Batch-to-Batch Consistency | Antibody density CV <15%; binding capacity CV <12%; elution >85% all lots; leaching <0.1%; each lot functionally validated |
For research use only, not for clinical use.
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