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Immunoprecipitation using Dynabeads® Protein A or G



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By popular demand, the NEW Immunoprecipitation Kits are now available! 
The kits include the required buffers, we have also improved the bead formulation.

This results in:

Reduced protocol time now <30 minutes!
Background is nearly completely eliminated.

Selection Guide
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How to avoid 2 typical IP problems


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Are you still using Sepharose®/Agarose slurry?



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.

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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






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)

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Other Product Advantages for Dynabeads® Protein A and Protein G:


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 stressStarting 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

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Shorter protocol time. Better yield and reproducibility with Dynabeads Immunoprecipitation Kit




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More information about Immunoprecipitation:


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 2 - Different elution properties for captured proteins

 

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.

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Table 1 - Properties of the recombinant Protein A and Protein G in Dynabeads® 


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 beads
640 µg mouse
IgG / ml beads
Optimal binding pH 8.2 7.0