Dynabeads™ Protein G - for oem / Industrial use only

Dynabeads™ with protein G coupled to their surface allow for small-scale purification of most mammalian immunoglobulins (Ig). The amount ofRead more

Catalog Number	Quantity
10005	50 mL

Catalog number 10005

Price (EUR)

Quantity:

50 mL

Dynabeads™ with protein G coupled to their surface allow for small-scale purification of most mammalian immunoglobulins (Ig). The amount of Ig captured is dependent on the concentration of Ig in the starting sample and the type and source of the Ig.

Dynabeads™ Protein G is widely used for small scale IgG purification and Ab labeling, immunoprecipitation (IP), chromatin immunoprecipitation (ChIP) and protein isolations. Magnetic separation technology is fast and gentle, causing minimal physical stress to your target proteins. This permits the isolation and concentration of labile composites that might otherwise dissociate or be damaged by proteases during long incubation times. Native protein conformation and intact, large protein complexes will be preserved. The product holds reputable Dynal high standards with respect to reproducibility and automation ability, and drives reliability for your assays.

Benefits and Features:

The gentle tube-based magnetic technology allows you to isolate intact proteins and protein complexes
The recombinant protein covalently coupled to the surface does not contain albumin-binding sites, thus avoiding background caused by co-purification of contaminating proteins.
Requires no columns, centrifugation steps or time-consuming pre-treatment of your samples. Scalable and automation-friendly magnetic technology.

Applications:
Small scale IgG purification, immunoprecipitation (IP), chromatin immunoprecipitation (ChIP) and protein isolations.

Binding capacity:
The amount of Ig captured is dependent on the concentration of Ig and Dynabeads™ Protein G in the starting sample. As a guideline 50 μl of Dynabeads™ Protein G will typically isolate 20 μg human IgG from a 100 μl sample (total volume) containing 100 μg (1mg⁄ml) human IgG. Predominant Fc binding allows optimal Ig orientation

Additional Info:
This specific product format is for large volume customers, available on an OEM basis. The product is also available in smaller volumes for end-users (Cat. no 100-03D and 100-04D). For further information and price quotes, please contact us at:ivd@invitrogen.com

For research use only. Not for use in diagnostic procedures.

Specifications

Concentration30 mg/mL

FormatBeads in suspension

High-throughput CompatibilityHigh-throughput Compatible

Ligand TypeProtein G

Product LineDYNAL, Dynabeads

Quantity50 mL

Diameter (Metric)2.8 μm

For Use With (Application)Immunoprecipitation

Product TypeProtein G Bead

Unit SizeEach

Contents & Storage

Store at 2°C to 8°C.

Frequently asked questions (FAQs)

What should I do if I get low binding to Dynabeads Protein A or Protein G magnetic beads?

Here are a few suggestions to try if you are getting low binding to the Dynabeads protein A/G beads:

-Verify the binding/specificity of your antibody to your antigen, e.g., by ELISA
-Check the binding of your antibodies to the beads; if the antibodies are not captured and bound to the beads, the immunoprecipitation experiment will not work
-If you have used the indirect IP method, try the direct IP method; conversely, if you have used the direct IP method, try the indirect IP method
-Check the amount of beads and sample volume with respect to the capacity of the different beads provided in the package inserts, and increase the amount of beads or the concentration of your antibody during coupling
-Increase the incubation time
-If using a commercially sourced antibody, confirm it is validated for IP
-Try another antibody

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What should I do when the antibodies coupled to Dynabeads Protein A or Protein G magnetic beads have lost function after crosslinking?

Here are a few suggestions you can try:

-Perform the IP without crosslinking the antibody to the beads
-Reduce the amount of crosslinker used to covalently attach the antibody to the beads
-Try a different crosslinker
-To prevent co-elution of antibody, try one of our surface-activated Dynabeads magnetic beads; this allows you to conjugate the antibody to the beads directly, through covalent binding

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

What should I do if Protein G is coming off the Dynabeads Protein G magnetic beads?

Here are some suggestions:

-Wash the beads prior to use in 100 mM glycine at a pH of 11.3 followed by a wash with 200 mM glycine at a pH of 2.8 for a very short period. Immediately transfer the beads to PBS with 0.01% Tween-20.
-Perform the elution in the low pH, glycine-based elution buffer recommended in the manual.
-Crosslinking of the antibody to the Protein G also crosslinks the Protein G to itself, which anchors it to the bead surface.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

How can I reduce background using Dynabeads Protein A or Protein G magnetic beads?

Protein G is coated onto hydrophilic beads. If your background is protein-mediated, then we normally suggest having a combination of blocking protein and non-ionic detergent both in the coupling and washing buffer to reduce nonspecific binding

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I was able to immunoprecipitate my protein using Dynabeads magnetic beads before, but after crosslinking the antibody to the beads I get no protein bands on my gel. What could be the problem?

The binding sites of your antibody have likely been altered by the crosslinking. When this occurs, your antibody will show reduced affinity or no affinity to its target antigen. Another consequence of crosslinking can also be increased affinity for unintended (nonspecific) targets. This is always a high risk with crosslinking, and it is a problem easily avoided by choosing another path to covalent antibody coupling. You can try using the Dynabeads Antibody Coupling Kit. This kit is a far superior solution for covalent antibody coupling to Dynabeads magnetic beads (compared to crosslinking with Dynabeads Protein A or G magnetic beads). The Dynabeads Antibody Coupling Kit is compatible with almost any antibody. It is designed specifically for covalent antibody coupling to Dynabeads magnetic beads. Unlike crosslinking, the Dynabeads Antibody Coupling Kit will not alter antibody specificity or affinity.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

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