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!)