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| Product Name | Anti-HA Magnetic Beads, 2 µm, High Affinity Clone 12CA5 |
| Catalog No. | MAGBEA-0030 |
| Description | Anti-HA magnetic beads with covalently immobilized high-affinity monoclonal anti-HA antibody (clone 12CA5, mouse IgG2b) for immunoprecipitation and purification of HA-tagged recombinant proteins. The 12CA5 clone is the original and most widely cited anti-HA antibody, recognizing the influenza hemagglutinin epitope YPYDVPDYA with an equilibrium dissociation constant in the sub-nanomolar range. The 2 µm superparamagnetic beads provide rapid magnetic separation kinetics and high binding capacity, making them suitable for quantitative immunoprecipitation, co-immunoprecipitation for protein interaction discovery, and tandem affinity purification (TAP) when combined with other epitope tag systems. |
| Intended Use | Immunoprecipitation and pull-down of HA-tagged recombinant proteins from mammalian, bacterial, yeast, and insect expression systems; Co-IP for protein interaction mapping; chromatin immunoprecipitation (ChIP) of HA-tagged chromatin-associated proteins; tandem affinity purification. |
| Principle / Technology | Monoclonal anti-HA (12CA5) antibody covalently coupled to magnetic bead surface recognizes HA epitope (YPYDVPDYA) at N-terminal, C-terminal, or internal positions with high affinity (Kd <1 nM); magnetic separation enables rapid washing and competitive peptide elution for gentle recovery of native protein complexes. |
| Detection Method | Equilibrate beads in lysis buffer; add to protein lysate (10-25 µL settled beads per 0.5-2 mg total protein); incubate 1-2 hours at 2-8 °C or 30-60 min at RT with rotation; wash 3-5×; elute with HA peptide (0.2-0.5 mg/mL) or 0.1 M glycine pH 2.8; or boil in SDS loading buffer for direct analysis. |
| Sample Type | Mammalian cell lysates (HEK293, HeLa, COS-7); bacterial lysates (E. coli); yeast lysates (S. cerevisiae, P. pastoris); baculovirus-infected insect cell lysates. |
| Performance Range / Specifications | Binding capacity: ≥500 µg HA-tagged protein per mL settled beads (protein-dependent); typical immunoprecipitation efficiency >80% of tagged protein from input lysate. |
| Sensitivity / LOD | Detection of HA-tagged proteins at endogenous expression levels (after stable integration); detectable by western blot from lysate equivalent of 1×10^5 cells. |
| Specificity | Clone 12CA5 specifically recognizes the HA nonapeptide sequence (YPYDVPDYA); minimal cross-reactivity with cellular proteins in mammalian lysates; does not recognize FLAG, Myc, V5, or GST epitope tags. |
| Reaction Conditions / Protocol | Bind at 2-8 °C for 1-2 hours; HA peptide elution at RT for 15-30 min; gentle competitive elution preserves protein complex integrity. |
| Components / Formulation | Anti-HA magnetic bead suspension (50% slurry in PBS, 0.1% BSA, 0.02% NaN3), HA peptide (YPYDVPDYA, lyophilized, 5 mg), 10× TBS buffer, protocol. |
| Storage Conditions | Store at 2-8 °C; do not freeze; HA peptide at -20 °C desiccated. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 0.5 mL settled beads (1 mL 50% slurry), 1 mL, 5 mL. |
| Product Form | Dark brown to black magnetic bead suspension; settles within 30 seconds in magnetic field. |
| Quality Control | Each lot tested for HA peptide binding capacity, specificity (HA-tagged vs. untagged protein enrichment >50:1), and absence of free antibody in supernatant. |
| Key Features | Original 12CA5 clone antibody; sub-nanomolar affinity for HA tag; 2 µm uniform beads for rapid separation; HA peptide elution included; low nonspecific binding; compatible with mammalian, bacterial, and yeast lysates. |
| Purity | Monoclonal antibody immobilized; no leaching detected by ELISA of supernatant; BSA carrier protein in storage buffer. |
| Concentration | 50% slurry; binding capacity ≥500 µg HA fusion protein per mL settled beads. |
| Activity / Unit Definition | Anti-HA 12CA5 Kd <1 nM for HA epitope (surface plasmon resonance); binding half-life >30 min. |
| Molecular Weight | Anti-HA 12CA5: ~150 kDa (mouse IgG2b). |
| Source / Origin | Mouse hybridoma clone 12CA5; magnetic beads synthesized from iron oxide core with hydrophilic polymer shell. |
| pH Range / Optimal pH | Optimal binding pH 7.0-7.5; compatible pH range 6.5-8.5. |
| Shipping Conditions | Cold pack (2-8 °C); do not freeze. |
| Expiration Date / Stability | 12 months at 2-8 °C; beads damaged by freezing — do not freeze. |
| Regulatory / Compliance | For research use only; contains NaN3 preservative. |
| Compatibility | Compatible with standard IP lysis buffers: 50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton X-100 or NP-40. Avoid >0.2% SDS. Reducing agents up to 5 mM DTT compatible. Magnetic separation compatible with standard magnetic racks for microcentrifuge tubes. |
| Recommended Buffer System | TBS or PBS with nonionic detergent and protease inhibitors, pH 7.4. |
| Application Notes / Precautions | Pre-clear lysate with empty beads (no antibody) or nonspecific IgG beads for 30-60 min before IP to reduce background. For Co-IP, use minimal detergent (0.1% NP-40) to preserve weak interactions. HA peptide elution is recommended for downstream functional assays as it preserves native protein conformation. For SDS-PAGE only, boiling beads directly in loading buffer is faster and gives complete recovery. Reuse beads up to 3 times after stripping with 0.1 M glycine pH 2.8. |
| Batch-to-Batch Consistency | Binding capacity within ±20% of reference lot; antibody activity verified per lot by immunoprecipitation of HA-tagged GFP standard. |
For research use only, not for clinical use.
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