Thermo Scientific™ Pierce™ DSG, No-Weigh™ Format

Product Code.	Quantity	unitSize
15859614	10 x 1 mg	Pack of 10
11836794	50 mg	50mg

Product Code. 15859614 Supplier Thermo Scientific™ Supplier No. A35392

Description

Thermo Scientific™ Pierce DSG is disuccinimidyl glutarate, a simple homobifunctional crosslinker based on the amine-reactive N-hydroxysuccinimide (NHS) ester group.

Thermo Scientific No-Weigh products are specialty reagents provided in a pre-aliquoted format. The pre-weighed packaging helps prevent loss of reagent reactivity and contamination over time by minimizing repetitive opening and closing of vial. The format enables use of fresh vial of reagent each time, eliminating the hassle of weighing small amounts of reagents and reducing concerns over reagent stability.

• Reactive groups: NHS ester (both ends)
• Reactive towards: amino groups (primary amines)
• Non-cleavable
• Water-insoluble (dissolve first in DMF or DMSO)
• Membrane-permeable, allowing for intracellular crosslinking
• Can increase crosslinking efficiency compared to DSS in some applications

Specifications

• Chemical Reactivity: Amine-Amine
• Cleavable: No
• Molecular Weight (g/mol): 326.26
• PEGylated: No
• Spacer Arm Length: 7.7 Å
• Content And Storage: Upon receipt store desiccated at 2 to 8°C.
• Cell Permeability: Yes
• Shipping Condition: Wet Ice
• Product Line: No-Weigh, Pierce
• Labeling Method: Chemical Labeling
• Crosslinker Type: Homobifunctional
• Reactive Moiety: NHS Ester
• Spacer: Short (<10 Å)
• Form: Powder
• Quantity: 10 x 1 mg
• Solubility: DMF, DMSO
• Format: Standard, Single-use
• Water Soluble: No
• Product Type: Crosslinker

Frequently Asked Questions (FAQs)

Can you provide the shelf-life for DSG (disuccinimidyl glutarate)?

DSG (disuccinimidyl glutarate) is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.

How should I dissolve and store DSG, No-Weigh Format?

DSG is an NHS-ester that hydrolyzes easily and is moisture-sensitive. For optimal results, we recommend storing DSG powder desiccated at 4 degrees C. To avoid moisture condensation onto the product, the vial must be equilibrated to room temperature before opening. We recommend resuspending DSG in DMSO, Anhydrous (Cat. No. D12345) right before use and not storing the solution.

How does DSP work as a crosslinker?

DSP is a homobifunctional, water soluble, cleavable, amine-reactive cross-linker. The amine-reactive ends are NHS-esters. This group reacts with primary amines, in proteins that translates to lysine groups and N-terminus amines. It reacts best in the pH range of 7.5 to 8.5, this ensures the amines are deprotonated and it limits the hydrolysis of the ester at higher pHs. NHS-esters are best used in a heavy molar excess over the number of amines present. This helps to overcome the competing hydrolysis of the ester to an inactive form.

How is the DSP cross-linking reaction quenched?

1M Tris (or glycine), pH 7.5 can be added to the reaction mixture to a final concentration of approximately 20-50 mM.

How is DSP cleaved?

The disulfIde bond in DSP's 12Å spacer arm can be cleaved by at least three different methods. The first is by incubating the crosslinker in 10-50 mM DTT for 30 minutes at 37 degrees C. Secondly, the disulfIde can be reduced by incubation in 5% 2-Mercaptoethanol (2-ME) in SDS-PAGE sample buffer at 100 degrees C for 5 minutes. Finally, the “-S-S-” bond can be cleaved by incubation in 100 mM sodium borohydride at room temperature for 30-60 minutes. Other reducing agents such as TCEP-HCl or 2-Mercaptoethylamine (2-MEA) should also work, however these applications have not been referenced.

Can stock solutions of DSP (Lomant's Reagent) be made?

Unfortunately not, the NHS-esters are unstable in aqueous solutions. They will begin to hydrolyze immediately upon contact with water. They cannot be kept in DMSO because it is hygroscopic and will draw moisture out of the air. NHS-crosslinking reagents work best in the pH range of 7.5 to 8.5. Within this range, the half-life of hydrolysis varies from I hour to 15 minutes, so NHS-crosslinking reagents work best when made up fresh.

Will the DSP (Lomant's Reagent) cross-linker penetrate a cell membrane?

Yes, it will penetrate a cell membrane.

What does "store under nitrogen" mean for DSP (Lomant's Reagent)?

A gentle stream of nitrogen gas is held over the opening of the vial to flush the oxygen from the vial. The cap is then immediately placed on the vial. If storing under nitrogen, caution should be taken to ensure that the stream of nitrogen is gentle enough, so that the powdered product will not be blown out of the bottle. If nitrogen is not available, weigh the material as quickly as possible and immediately cap. Store desiccated at 4 degrees C.

What are the recommended storage conditions for DSP (Lomant's Reagent)?

The product is shipped ambient. Upon receipt. the product should be stored at 4 degrees C in a dessicator or with a desiccant bag, and the vial should be warmed to room temperature before opening.

What is the expiration date for DSP (Lomant's Reagent)?

This product has no set expiration date. LSG/Rockford, IL had monitored the bulk material in such a manner that allows us to warrant the product for one year from the date of shipment. This product may still be usable beyond the one year if handled properly. However, using material older than one year is typically not recommended since the material easily hydrolyzes upon exposure to moisture.

How much DSP (Lomant's Reagent) should be used for cross-linking?

A 10 fold molar excess of cross-linker to protein is recommended for protein solutions greater than 5 mg/mL. A 20-50 fold molar excess of cross-linker to protein is recommended for protein solutions less than 5 mg/mL.

Can the DSP cross-linking agent be added directly to the reaction mixture?

No. It must first be dissolved in dry DMSO or DMF.

What is the recommended concentration of the DSP (Lomant's Reagent) stock solution?

An initial stock concentration of 5-10 mM (~3-6 mg/mL) is recommended.

Which components are not recommended in buffers to be used with DSP?

Buffers containing primary amines, Tris, ethanolamine, glycine, or lysine should not be used, since they will quench the reaction. Imidazole and gluathione are also not recommended as buffer components.

What is the recommended sample buffer and reconstitution buffer to obtain optimal cross-linking using DSP (Lomant's Reagent)?

DSP is first dissolved in an organic solvent (such as dry DMSO) and added to an aqueous reaction mixture such as 20 mM sodium phosphate, 0.15 M NaCl, pH 7.5 (PBS). HEPES, bicarbonate/carbonate, and borate buffers may be substituted for PBS. Other buffers may also be substituted provided they do not contain primary amines. Recommended pH is 7-8. Once the cross-linker is reconstituted, use immediately.

Is DSP (Lomant's Reagent) water soluble?

No, the reagent must first be reconstituted utilizing an organic solvent, such as dry DMSO or DMF.
Note: It is recommended that the DMSO/organic solvent should not exceed 10% of the final reaction volume.

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