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How-to-Choose-the-Right-Immunoprecipitation-Product-for-Your-Experiment

How to Choose the Right Immunoprecipitation Product for Your Experiment

richardjgray
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How-to-Choose-the-Right-Immunoprecipitation-Product-for-Your-Experiment

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  1. How to Choose the Right Immunoprecipitation Product for Your Experiment CD Magnetic Beads vs. Agarose Resin Capacity Magnetic beads If the sample size is < 2 mL, magnetic beads are best for manual and automated standard IP, Co-IP, ChIP, ChIP-Seq, RIP, and pull-down reactions for immediate assay analysis. Yield Reproducibility Purity-Specificity Agarose Resin Magnetic Beads Agarose resin Ease of Use If the sample size is > 2 mL and the goal is aiming to purify a sufficient amount of target protein for multiple downstream assays, agarose is more appropriate for column affinity chromatography or individual large scale spin cup IP reactions. Speed Multiple Samples Automation Tel: 1-631-624-4882 Email: info@cd-bioparticles.com

  2. www.cdparticles.com 1 2 Starting with: Absolute Mag™ Protein A or Absolute Mag™ Protein G 3 Absolute Mag™ Beads Protein A or G Antibody 4 Target Protein Nonspecific Protein IP with Magnetic Beads 2. Add sample to the beads 1. Binding of the antibody to the beads 4. Elute the protein 3. Wash the protein-bead complex Advantages of Magnetic Beads for IP Magnetic beads are a faster, simpler, and more efficient method than the use of traditional agarose or other agarose resins for target protein pull-down, and have become the gold standard for IP and pull-down analysis. Absolute Mag™ magnetic beads help you achieve the best balance between high yield, high reproducibility and low non-specific binding, low cost. Tel: 1-631-624-4882 Email: info@cd-bioparticles.com

  3. www.cdparticles.com Capacity and yield Magnetic beads are nonporous with a smooth outer surface. Although they are much smaller than agarose beads, they have a lower theoretical binding capacity than agarose. But, when individual antibodies are immobilized on sponge-like agarose, these antibodies are not always accessible to the target proteins in a sample, which results in the antibody lost during the washing steps. On the contrary, they are all accessible to an antigen if antibodies are bound to the surface of magnetic beads, and there is little chance for antibodies to be lost during the gentle washing steps. Fast & Easy Highly Sensitive Flexible Highly Reproducible Automation of IP Protocol Low Antibody Savings Background Benefits for Absolute Mag™ Technology Reproducibility and purity When using magnetic beads and a magnet, all the beads are held firmly to the side of the tube, and thus all buffers are able to be removed without touching the bead-pellet. Moreover, magnetic beads are uniform in size and all interactions occur on their outer smooth surface, so they can usually provide higher reproducibility and purity than agarose. Pre-clearing is generally not necessary with magnetic beads. Ease of use, speed, and automation An individual magnetic bead IP experiment can be completed within 30 min and can process multiple samples easily and rapidly. In addition, small benchtop instrumentation is available for the automation of the whole IP protocols using magnetic beads. Tel: 1-631-624-4882 Email: info@cd-bioparticles.com

  4. www.cdparticles.com Choose the Right Immunoprecipitation Product for Your Experiment Antibody-binding IP products If you have an antibody that recognizes your target protein (or your downstream assay is mass spectrometry, or you don’t want the antibody co-eluted with your target protein), you can choose antibody-binding products. Secondary Antibodies Surface-activated Beads Protein A, G Binding properties Non-covalent Covalent Non-covalent Antibody co-eluted off the beads Yes No Yes Different ligands bind different species and antibody subclasses with different specificity IgGs Type of ligand All antibodies Mass spec compatible No Yes Not validated Nonspecific binding Low Low Ultra-low Products Absolute Mag™ Amine Absolute Mag™ Carboxyl Absolute Mag™ Epoxy Absolute Mag™ Tosyl more Absolute Mag™ Protein A Absolute Mag™ Protein G Absolute Mag™ Anti-Goat IgG Absolute Mag™ Anti-Rabbit IgG Absolute Mag™ Anti-Mouse IgG Absolute Mag™ Anti-Rat IgG Absolute Mag™ Anti-Human IgG Tel: 1-631-624-4882 Email: info@cd-bioparticles.com

  5. www.cdparticles.com Biotin-binding IP products If you have a sample rich in soluble IgGs If you have a recombinant antibody lacking Fc regions Absolute Mag™ Streptavidin Magnetic Particles If you already have streptavidin coated magnetic beads in the If you need a bead compatible with mass spectrometry Recombinant protein (fusion tag) IP products Absolute Mag™ Ni-NTA Absolute Mag™ Glutathione If you have a recombinant (fusion tagged) protein Absolute Mag™ Anti-HA Absolute Mag™ Ni-IDA For more information, view our website: www.cd-bioparticles.com Email: info@cd-bioparticles.com Tel: 1-631-624-4882 Fax: 1-631-938-8221 Address: 45-1 Ramsey Road, Shirley, NY 11967, USA 5

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