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  6. Thermo Scientific™ Detoxi-Gel™ Endotoxin Removing Gel

Thermo Scientific™ Detoxi-Gel™ Endotoxin Removing Gel

Catalog No. PI20339
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PI20339 10 mL
PI20340 1000 mL
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Catalog No. PI20339 Supplier Thermo Scientific™ Supplier No. 20339
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Description

Bind and extract endotoxins (lipopolysaccharides or LPS) from antibody or protein samples using bulk resin or columns of immobilized Polymixin B ligand.

Thermo Scientific™ Detoxi-Gel Endotoxin Removing Gel and Columns provide a quick method to remove endotoxins (lipopolysaccharides or LPS) from samples.

Detoxi-Gel Resin is a beaded agarose support onto which polymixin B has been immobilized. This ligand binds to the lipid A portion of lipopolysaccharides (LPS), which are the dominant form of endotoxins that affect biological samples. Even low levels of endotoxins can be toxic to cells or organisms and must be removed before biological samples can be added to cells for various treatment experiments. Detoxi-Gel Resin provides a quick method for removal of endotoxins from samples. It is available in convenient pre-packed columns and two sizes of slurry for use in small-scale or large-scale processes.

Highlights:

  • Affinity-based – uses immobilized polymixin B to remove endotoxins by binding to their lipid A domains

  • High capacity – 1mL of Detoxi-Gel Resin removes at least 9995 endotoxin units from a 5mL sample containing 10,000 EU of lipopolysaccharide (LPS)

  • Reusable – no loss in binding capacity even after 10 regenerations; requires activation and regeneration with 1% sodium deoxycholate (Part No. 89904)

  • Testing required – protein recovery is dependent upon protein type and concentration; empirical testing is required to validate the method for particular protein samples

  • Versatile – remove endotoxin from protein solutions, cell culture media and pharmacological components

Requires:

Sodium deoxycholate (Part No. 89904) for resin activation and regeneration.

Specifications

Capacity (Metric) 4000 to 6000 EU Binding per mL Gel
Content And Storage Store at 4°C.
Form Liquid
Product Line Detoxi-Gel
Quantity 10 mL
Target Endotoxin Removal
Type Endotoxin Removing Gel
Stationary Phase Polymixin B
Sufficient For 1 mL of gel binds 4000-6000 EU
Format Bottle
Column Type Affinity, Agarose Resin
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Frequently Asked Questions (FAQs)

Why must I regenerate the Detoxi-Gel Endotoxin Removing Gel column before and after each use?

It is possible that the column may have been contaminated with endotoxin during storage and handling. Equilibrating the column with 1% sodium deoxycholate followed by pyrogen-free washing ensures that the column is free of endotoxin before the sample is added. The column must be regenerated following endotoxin removal from a sample to make sure the endotoxin that has bound to the column from the sample is washed off.

What is the void volume of the Detoxi-Gel Endotoxin Removing Gel column?

The Thermo Scientific 1 mL Detoxi-Gel Column has a void volume of 0.3-0.5 mL. This is the volume of solution that must be collected from the column before the sample will begin to come off the column.

What is the protocol for using Detoxi-Gel Endotoxin Removing Gel?

1. Equilibrate the column with 5 column volumes of 1% sodium deoxycholate in pyrogen-free water, followed by 3 column volumes of pyrogen-free water or buffer of choice.
2. Apply sample, being sure not to exceed the binding capacity of the gel.
3. Collect sample by gravity flow by applying additional pyrogen-free water or buffer.
4. Regenerate the column with 5 column volumes of 1% sodium deoxycholate in pyrogen-free water, followed by 3 column volumes of pyrogen-free water.

What is the binding capacity of Detoxi-Gel Endotoxin Removing Gel?

Detoxi-Gel Support has the capacity to bind up to 2µg LPS/mL of gel. Because one endotoxin unit (EU) is approximately 0.1 ng/mL, the gel has the potential for binding up to 20,000 EU/mL of gel.

What is Detoxi-Gel Endotoxin Removing Gel, and how does it work?

Detoxi-Gel Support consists of 6% cross-linked beaded agarose that has Polymixin B immobilized onto it. Polymixin B is an antibiotic that contains a cationic cyclopeptide with a fatty acid chain that can neutralize the biological activity of endotoxins by binding to the lipid A portion of the bacterial lipopolysacharide (LPS). After the endotoxin from a sample binds to the immobilized Polymixin B, the sample can be collected and the endotoxin can be eluted using 1% sodium deoxycholate.

Is it possible for substances other than LPS to bind to the Detoxi-Gel Support?

Yes. Hepatitis B surface antigen, for example, is known to stick to the column. Adding 0.5 M NaCl to the sample and elution buffers can sometimes break the affinity for certain substances that bind nonspecifically, releasing them from the gel.

How many times can the same Detoxi-Gel Endotoxin Removing Gel be used?

The columns may be reused at least 10 times without loss of activity.

Do any substances interfere with the binding of LPS to Detoxi-Gel Support?

Yes. Other detergents and high levels of chaotropes (urea and guanidine) can reduce the affinity of the Polymixin B for the LPS. Proteins such as BSA can bind tightly to endotoxin, reducing its ability to interact with and bind to the Detoxi-Gel Support. This reduction in binding capacity can sometimes be overcome by increasing the volume of Detoxi-Gel Support to LPS. However, some proteins bind tightly to endotoxin without inhibiting its binding to the support. In this case, the protein will remain bound to the gel with the endotoxin, possibly causing a loss of the protein of interest.
Can the Detoxi-Gel Endotoxin Removing Gel (Cat. No. 20339) be used to remove LPS from nucleic acids?
Yes. Detoxi-Gel Endotoxin Removing Gel (Cat, No. 20339) is compatible with nucleic acids and proteins. We recommend slowly pumping the sample over the gel for extended periods of time instead of using a single gravity filtration through the column.

For Research Use Only. Not for use in diagnostic procedures.