Protein A、G、L functional magnetic microparticles (PuriMag G- ProA、ProG、ProL Beads)
- BrandPuriMag
- TypePuriMag G Series
|
Ordering information |
|||
|
Name |
Cat. No. |
Vol. |
Scheme |
|
G-ProA |
PMG013-2 |
2 ml |
|
|
G-ProG |
PMG014-2 |
2 ml |
|
|
G-ProA/G |
PMG029-2 |
2 ml |
|
|
G-ProL |
PMG030-2 |
2 ml |
|
1. Overview
PuriMag? Beads enable rapid magnetic separation of antibodies through affinity binding. Recombinant Protein A, G, or L covalently coupled to surface-blocked beads binds antibodies from diverse species, facilitating purification from crude extracts. Immunoprecipitation assays using these protein-coated beads deliver low background and high target antigen yield. Crosslinker chemistry allows permanent antibody immobilization on magnetic particles, preventing IgG leakage in IP/Co-IP experiments.
Core Applications
Antibody isolation from serum, cell culture supernatants, or ascites fluid
Immunoprecipitation (IP) and co-immunoprecipitation (Co-IP) of antigens from cell/tissue extracts
Protein L-Specific Features:
Selectively binds mouse/human antibodies via kappa light chains
Does not bind bovine IgG, making it ideal for purifying monoclonal antibodies from bovine serum-supplemented cell cultures
Standard Workflows
(Recover purified solutions using magnetic racks or automated instruments)
Product Specifications
Unified Advantages Across Product Line
Antibody Purification: Incubate beads with antibody solution → magnetic separation
Immunoprecipitation: Add beads to antigen samples containing antibodies → form antibody-antigen complexes → elute targets
PuriMag? G-ProA Beads
Immobilized protein: Recombinant Protein A (44.7 kDa; apparent MW ~47 kDa by SDS-PAGE)
Structural features: 5 Fc-binding domains per protein
Species coverage: Mouse, Human, Rabbit, Pig, Dog, Cat
Surface treatment: Proprietary blocking minimizes non-specific binding in complex biological samples
PuriMag? G-ProG Beads
Immobilized protein: Recombinant Protein G (21.6 kDa; apparent MW ~32 kDa by SDS-PAGE)
Engineering optimizations:
2 Fc-binding domains per protein
Albumin/cell-binding domains removed to reduce background
Species coverage: Mouse, Human, Rabbit, Cow, Goat, Sheep
Performance assurance: Includes optimized buffer components and detailed protocols
PuriMag? G-ProA/G Beads
Composition: 1:1 physical blend of G-ProA and G-ProG beads
Key advantage: Combines binding specificities of both proteins
PuriMag? G-ProL Beads
Immobilized protein: Recombinant Protein L (35.8 kDa; apparent MW ~36 kDa by SDS-PAGE)
Binding mechanism:
4 immunoglobulin-binding domains per protein
Targets IgG/IgM/IgA/IgE/IgD, scFv, and Fab fragments via kappa light chains
Unique value: Bovine IgG non-reactivity for serum-supplemented culture systems
High efficiency: Delivers IP target antigen yields comparable or superior to competitors
Ultra-low non-specific binding: Pre-blocked surfaces ensure contaminant-free eluates (e.g., no residual proteins from complex lysates)
Operational consistency: Magnetic separation eliminates resin loss associated with centrifugation
Workflow versatility: Compatible with both manual and automated high-throughput platforms
2. product description
Product Specifications
Description
Polymer coated Fe3O4 nanoparticles
Particle Size
200 nm
Number of Beads
~1.7×1010 beads/mg
Matrix
Proprietary polymer
Functional group
ProA, ProG, ProL group
Binding capacity
>60 μg rabit IgG / mg Beads
Magnetization
60~70 EMU/g
Formulation
10mg/mL in DI water, 0.05% NaN3，0.01% Tween 20，0.05% BSA
Storage
1 year at 2~8 ℃. Do not freeze.
3. Instructions for Use
3.1 Precautions
Do not centrifuge, dry, or freeze PuriMag? G-ProA beads. These actions cause bead aggregation and loss of binding activity.
For optimal antibody binding and dispersion, include 0.025%–0.1% non-ionic detergent (e.g., Tween-20) or zwitterionic detergent (e.g., CHAPS) in the binding buffer, and mix beads during incubation.
To minimize protein degradation, add protease inhibitors when preparing cell lysates.
For single-use applications, low-pH elution is acceptable. Optimal elution time is ≤10 min; exceeding this may increase non-specific binding and reduce yield.
For downstream Western blotting with rabbit antibodies (primary/secondary): Elute in SDS-PAGE sample buffer at room temperature.
For all other antibody species: Boiling beads in SDS-PAGE sample buffer is acceptable for single-use applications.
Note: Boiling causes irreversible bead aggregation and loss of activity.
PuriMag? G-ProA beads are compatible with:
Small-scale antibody purification
Immunoprecipitation
Western blot analysis
Select the appropriate protein-coated beads when isolating polyclonal IgG from different serum sources.
3.2 Antibody Purification Protocol
A. Buffers
Binding/Wash Buffer: Tris-buffered saline (50 mM Tris, 150 mM NaCl, pH 7.6 adjusted with HCl) + 0.05% Tween-20.
Elution Buffer: IgG elution buffer or 0.1 M glycine, pH 2.0.
Neutralization Buffer: High-ionic strength alkaline buffer (e.g., 1 M phosphate or 1 M Tris, pH 7.5–9).
B. Antibody Purification (from Serum, Cell Culture Supernatant, or Ascites)
Note: Thoroughly resuspend beads by inversion, gentle vortexing, or rotation before use.
1.Transfer 50 μL (0.50 mg) beads to a 1.5 mL microcentrifuge tube. Add 150 μL Binding/Wash Buffer and mix gently by vortexing.
2.Place tube in magnetic stand. Collect beads against tube wall. Remove and discard supernatant.
3.Add 1 mL Binding/Wash Buffer. Invert tube or vortex gently for 1 min. Collect beads magnetically; discard supernatant.
4.Dilute 10 μL sample with 490 μL Binding/Wash Buffer (final volume: 500 μL).
If sample volume <500 μL, adjust to 500 μL with buffer.
5.Add diluted sample to pre-washed beads. Mix gently by vortexing/inversion.
6.Incubate with mixing at RT for 1 h.
7.Collect beads magnetically. Discard supernatant.
8.Wash beads:
Add 500 μL Binding/Wash Buffer, mix thoroughly.
Collect beads magnetically; discard supernatant.
Repeat twice (total 3 washes).
9.Add 100 μL Elution Buffer. Mix thoroughly and incubate at RT for 10 min with intermittent mixing.
10.Collect beads magnetically. Transfer supernatant (containing eluted antibodies) to a new tube.
Neutralize: Add 15 μL Neutralization Buffer per 100 μL eluate.
*Minimum recommended bead volume for antibody purification: 50 μL.*
3.3 Co-Immunoprecipitation Protocol
A. Buffers
Binding Buffer: Buffer used to prepare the antigen sample.
Wash Buffer: Tris-buffered saline (50 mM Tris, 150 mM NaCl, pH 7.6 adjusted with HCl) containing 0.05% Tween-20.
Elution Buffer: IgG elution buffer or 0.1 M glycine, pH 2.0.
Neutralization Buffer: 1 M Tris, pH 8.5.
B. Immunoprecipitation
Note: This protocol serves as a general guideline; optimize for each specific application.
1.Mix the antigen sample with 5–10 μg antibody. Adjust reaction volume to 500 μL using cell lysis buffer. Incubate with mixing:
1–2 h at RT or overnight at 4°C.
2.Transfer 25 μL (0.25 mg) Pierce Protein A Magnetic Beads to a 1.5 mL microcentrifuge tube.
3.Add 175 μL Wash Buffer to beads. Mix gently by vortexing.
4.Place tube in magnetic stand. Collect beads against tube wall. Discard supernatant.
5.Add 1 mL Wash Buffer. Invert tube or vortex gently for 1 min. Collect beads magnetically. Discard supernatant.
6.Add antigen/antibody mixture to pre-washed beads. Incubate with mixing at RT for 1 h.
7.Collect beads magnetically. Save the flow-through for analysis.
8.Wash beads:
Add 500 μL Wash Buffer, mix gently.
Collect beads magnetically; discard supernatant.
Repeat twice (total 3 washes).
9.Add 500 μL ultrapure water, mix gently. Collect beads magnetically; discard supernatant.
(For research use only!)
