Immunoprecipitation using Dynabeads® Protein A or G
Obs:. Envio de 1(um) DynaMag por cliente após preenchimento da pesquisa.
Cliente poderá se cadastrar e participar da promoção apenas uma vez.
Dúvidas envie um e-mail para: marketing.br@invitrogen.com
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
________________________________________________________________________________________________________________________
How to avoid 2 typical IP problems
________________________________________________________________________________________________________________________
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.
________________________________________________________________________________________________________________________
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)
________________________________________________________________________________________________________________________
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
________________________________________________________________________________________________________________________
Shorter protocol time. Better yield and reproducibility with Dynabeads Immunoprecipitation Kit
________________________________________________________________________________________________________________________
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.
________________________________________________________________________________________________________________________
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 |
