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| Product Name | Anti-FLAG M2 Magnetic Beads, 2 µm, Monoclonal Antibody, Mouse IgG1 |
| Catalog No. | MAGBEA-0029 |
| Description | Anti-FLAG M2 magnetic beads with covalently immobilized monoclonal anti-FLAG M2 antibody (mouse IgG1, clone M2) for rapid and efficient immunoprecipitation (IP), co-immunoprecipitation (Co-IP), and pull-down of FLAG-tagged recombinant proteins from cell lysates and in vitro translation systems. The M2 clone recognizes the FLAG epitope (DYKDDDDK) at N-terminal, C-terminal, or internal positions, and binding is calcium-independent (unlike M1 clone). The 2 µm superparamagnetic iron oxide particles encapsulated in a hydrophilic polymer shell provide high surface area for antibody conjugation and rapid magnetic separation (<30 seconds). |
| Intended Use | Immunoprecipitation and affinity purification of FLAG-tagged recombinant proteins; co-immunoprecipitation for protein-protein interaction analysis; pull-down of FLAG-tagged protein complexes; chromatin immunoprecipitation (ChIP) with FLAG-tagged transcription factors. |
| Principle / Technology | Monoclonal anti-FLAG M2 antibody (clone M2) covalently immobilized on magnetic bead surface; FLAG epitope (DYKDDDDK) specifically recognized with dissociation constant Kd ~1 nM; calcium-independent binding enables use with EDTA-containing buffers; magnetic separation enables rapid washing and elution. |
| Detection Method | Add magnetic beads to cell lysate (10-25 µL settled beads per 0.5-1 mg total protein); incubate with end-over-end rotation at 2-8 °C for 1-2 hours or room temperature for 30 min; magnetically separate; wash 3-5× with lysis/wash buffer; elute bound protein with 3× FLAG peptide (0.1-0.5 mg/mL) or low pH glycine; or boil beads in SDS loading buffer for direct SDS-PAGE analysis. |
| Sample Type | Mammalian cell lysates, bacterial lysates, yeast lysates, insect cell lysates, in vitro transcription/translation reactions expressing FLAG-tagged proteins. |
| Performance Range / Specifications | Binding capacity: ≥600 µg FLAG-BAP (bacterial alkaline phosphatase fusion) per mL settled beads; typical yield: 1-10 µg FLAG-tagged protein from 1 mg total cell lysate when expressed at moderate levels. |
| Sensitivity / LOD | Detection of FLAG-tagged proteins at expression levels as low as 0.01% of total cellular protein by western blot after IP; as few as 10 ng FLAG peptide detectable after enrichment. |
| Specificity | M2 monoclonal antibody specifically recognizes the FLAG octapeptide (DYKDDDDK); <0.1% cross-reactivity with 3× FLAG (which also contains DYKDHD epitope recognized by M2 at reduced affinity); no cross-reactivity with HA, Myc, GST, His6, or V5 epitope tags. |
| Reaction Conditions / Protocol | Bind at 2-8 °C for optimal specificity or RT for convenience; bind-reaction time 30 min to 2 hours; elute with 3× FLAG peptide (competitive elution, gentle) or 0.1 M glycine pH 3.0 (pH elution, dissociating). |
| Components / Formulation | Anti-FLAG M2 magnetic bead suspension (50% slurry in PBS with 0.1% BSA and 0.02% NaN3), 3× FLAG peptide (lyophilized, 5 mg), 10× TBS buffer, protocol. |
| Storage Conditions | Store bead suspension at 2-8 °C; do not freeze; FLAG peptide at -20 °C. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 0.5 mL settled beads (1 mL 50% slurry), 1 mL, 5 mL; 3× FLAG peptide: 5 mg. |
| Product Form | Dark brown to black suspension; beads settle rapidly in magnetic field. |
| Quality Control | Each lot tested for FLAG peptide binding capacity (≥600 µg FLAG-BAP/mL beads), specificity (compare binding of FLAG-tagged vs. untagged protein), and absence of detectable antibody leaching (by SDS-PAGE/silver stain of eluate without peptide). |
| Key Features | Monoclonal M2 anti-FLAG antibody; Ca2+-independent binding; 2 µm uniform particle size; rapid magnetic separation (<30 seconds); low non-specific binding; 3× FLAG peptide elution included; stable at 2-8 °C. |
| Purity | Monoclonal antibody covalently immobilized; no free antibody detected in supernatant; BSA carrier protein and NaN3 preservative in storage buffer. |
| Concentration | 50% slurry in PBS/BSA/NaN3; typical use: 25 µL slurry (12.5 µL settled beads) per IP. |
| Activity / Unit Definition | Binding capacity: ≥600 µg FLAG-BAP per mL settled beads; M2 Kd ~1 nM for FLAG octapeptide. |
| Molecular Weight | Anti-FLAG M2: ~150 kDa (monoclonal mouse IgG1). |
| Source / Origin | Monoclonal mouse anti-FLAG M2 antibody (clone M2) from mouse hybridoma; magnetic beads synthesized from iron oxide with hydrophilic polymer coating. |
| pH Range / Optimal pH | Binding and wash buffer pH 7.0-8.0; elution buffer (FLAG peptide): pH 7.4; low pH elution: 0.1 M glycine pH 3.0. |
| Shipping Conditions | Cold pack (2-8 °C); do not freeze beads. |
| Expiration Date / Stability | 12 months at 2-8 °C; do not freeze — freezing damages magnetic bead polymer coating. |
| Regulatory / Compliance | For research use only; not for diagnostic or therapeutic applications. Contains sodium azide (0.02%) as preservative — handle accordingly. |
| Compatibility | Compatible with most cell lysis buffers: RIPA (≤0.1% SDS), NP-40, Triton X-100, CHAPS. Avoid >0.5% SDS which may denature antibody. Compatible with reducing agents (DTT ≤5 mM, β-ME ≤1%). Protease and phosphatase inhibitors recommended. Magnetic separation compatible with standard magnetic racks for 1.5/2 mL tubes and 15/50 mL tubes. |
| Recommended Buffer System | TBS or PBS with 0.1% nonionic detergent, pH 7.4. |
| Application Notes / Precautions | Pre-wash beads with lysis buffer before adding to lysate. Do not vortex beads — mix by gentle inversion or end-over-end rotation. For Co-IP of weak interactions, use lower wash salt concentration (100-150 mM NaCl) and shorter wash times. For FLAG peptide elution, incubate with gentle shaking for 30 min at 2-8 °C for optimal recovery. Beads can be regenerated by stripping with 0.1 M glycine pH 3.0 and re-equilibrating in TBS; reuse up to 3-5 times for same protein target. |
| Batch-to-Batch Consistency | Binding capacity within ±20% of reference lot; no detectable antibody leaching; bead size distribution within specification (PDI <0.1). |
For research use only, not for clinical use.
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