Immunoprecipitation using Dynabeads® Protein A or G
How to avoid 2 typical IP problems
Are you still using Sepharose®/Agarose slurry?
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Dynabeads® Protein A and Protein G are the new gold standard for immunoprecipitation.
Magnetic handling is fast, efficient and extremely gentle on your target proteins. All steps take place in a single tube, with no centrifugations. Bound only by affinity interactions, your protein and protein complexes are preserved intact. Your immunoprecipitation is ensured maximum sensitivity and no loss of target protein.
Bring your research tools up to date now!
Sepharose® is a trademark of GE Healthcare companies.
Which benefit is most important for you?
Eliminate background: Less non-specific binding |
Reduced protocol time:30 minutes start to finish |
Easier handling:Single tube protocol |
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Why not get it all ! With Dynabeads® Protein A and Dynabeads® Protein G.
Take advantage of this introductory offer now!
(offer valid in Europe, US, Canada, Australia, New Zealand and Singapore)
Other Product Advantages for Dynabeads® Protein A and Protein G:
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- Perfect for small-scale IgG purification and immunoprecipitation
- No columns, centrifugations, or time-consuming pre-treatment of your samples
- Easy handling and simple protocol (workflow)
- Gentle, minimal physical stress
- Starting sample can be saliva, plasma, ascites, serum, and tissue culture or hybridoma supernatants
- The recombinant protein A/G on the beads contains no albumin binding sites, thus avoiding co-purification of contaminating proteins
Shorter protocol time. Better yield and reproducibility with Dynabeads Immunoprecipitation Kit
More information about Immunoprecipitation:
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Binding of antibodies to the beads in solution is an equilibrium reaction. To capture as many antibodies as possible, be aware of the following: The reaction volume should be kept low to maintain high concentrations of beads and antibody. There is no need for pre-treating the samples (even viscous samples), so no dilutions of sample or beads should be done. If the antibody concentration in the sample is suspected to be very low, the amount of beads can be increased.
Instead of eluting off the antibody, the Dynabeads® may be resuspended in a Na-phosphate buffer (Figure 2). The bead-bound antibody can be used for immunoprecipitation of specific proteins in any type of sample. To prevent co-elution of the antibody, it is possible to cross-link the antibody to protein A/G prior to the immunoprecipitation steps. This is not necessary for downstream SDS-PAGE followed by autoradiography or western blotting, and optional for Silver or Coomassie® staining.
Figure 1 - Affinity purification of human IgG and immunoprecipitation of human serum albumin from 2 µl serum, using Dynabeads® Protein A. Lane 1 - Purified IgG from serum. Lane 2: Immunoprecipitated human serum albumin (cross-linking omitted thus antibody present). Lane 3: 50 x diluted serum. Lane 4: LMW.
Figure 2 - Different elution properties for captured proteins 
Table 1 - Properties of the recombinant Protein A and Protein G in Dynabeads®
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Dynabeads® Protein A and Dynabeads® Protein G bind with different affinities depending on the immunoglobulin type and species. Binding occurs mainly through the Fc region. The binding capacity will depend on the initial concentration of antibodies in the sample and the source of the antibody. These data are based on isolation from a sample containing 100 µg immunoglobulin per ml.
Protein A Protein G Source Bacillus E.coli Molecular weight 45 kDa 17 kDa No.of binding sites for IgG 4 2 Binding capacity* 250 µg human
IgG / ml beads640 µg mouse
IgG / ml beadsOptimal binding pH 8.2 7.0
Related Info
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- Product Selection Guide
- It's time to update your immunprecipitation method: Brochure (pdf)
- Immunoprecipitation with Dynabeads® Protein A or Protein G: Application Note (pdf)
- References for Dynabeads® Protein A and Protein G
- Immunoprecipitation Crosslinking Protocol
- Protein Isolation: Brochure (pdf)
- Magnets
- Immunoprecipitation Kit
Protein A NEW! - immunoprecipitation Kit
Protein G NEW! - Dynabeads® Protein A
- Dynabeads® Protein G
Online catalog pages:
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