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  6. Thermo Scientific™ DSS (disuccinimidyl suberate), No-Weigh™ Format

Thermo Scientific™ DSS (disuccinimidyl suberate), No-Weigh™ Format

Product Code. 10138353 Shop All Thermo Scientific Products
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Product Code. 10138353 Supplier Thermo Scientific™ Supplier No. 21655

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Description

Crosslink amines to amines with disuccinimidyl suberate composed of NHS-ester groups with a membrane-permeable, mid-length spacer for intracellular protein crosslinking.

Thermo Scientific™ Pierce DSS, also called disuccinimidyl suberate, is noncleavable and membrane permeable crosslinker that contains an amine-reactive N-hydroxysuccinimide (NHS) ester at each end of an 8-carbon spacer arm.

Highlights:

  • Chemical crosslinking of intracellular proteins prior to cell lysis and immunoprecipitation
  • “Fix” protein interactions to allow identification of weak or transient protein interactions
  • Protein crosslinking to create bioconjugates via single-step reactions
  • Crosslink oriented antibodies to Protein A or G agarose
  • Immobilize proteins onto amine-coated surfaces
TRUSTED_SUSTAINABILITY

Specifications

Chemical Reactivity Amine-Amine
Cleavable No
Description DSS
Molecular Weight (g/mol) 368.35
PEGylated No
Spacer Arm Length 11.4 Å
Content And Storage Upon receipt store desiccated at 4°C.
Cell Permeability Yes
Shipping Condition Ambient
Product Line Pierce
Labeling Method Chemical Labeling
Crosslinker Type Homobifunctional
Reactive Moiety NHS Ester
Spacer Medium (10 to 30 Å)
Form Powder
Quantity 50 mg
Solubility DMF, DMSO
Format Standard, Single-use
Water Soluble No
Product Type Crosslinker
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Frequently Asked Questions (FAQs)

Will the DSS crosslinked product become unlinked as a result of boiling, incubation with SDS, high salt, high or low pH or any other means?

No. DSS crosslinks are amide bonds (same as peptide bonds).
What is the recommended buffer for optimal DSS crosslinking?

We recommend the following buffers: 20 mM sodium phosphate, 0.15 M NaCl, PBS, pH 7.5. Or HEPES, bicarbonate-carbonate and borate buffers may be substituted for PBS. Nearly any non-amine buffer pH 7-8 can be used.
What is the difference between DSS and BS3 crosslinkers?

BS3 is the water-soluble analogue of DSS (i.e., DSS is the membrane-permeable analog of BS3). DSS is water-insoluble and membrane-permeable, so it can be used for both intracellular and cell surface crosslinking. BS3 is water-soluble and membrane-impermeable, so it can be used for cell-surface crosslinking.
What buffers are not recommended for DSS crosslinking?

Any buffer containing primary amines (Tris, glycine, lysine, and ethanolamine) will not work. Glutathione- and imidazole-containing buffers are also not recommended.
What are the reactive groups in the DSS crosslinking reagent and what are they reactive toward?

The reactive groups are NHS esters that are reactive toward primary amines.
Is the DSS crosslinking reagent water-soluble?

No. The reagent must first be reconstituted in an organic solvent before 10-fold dilution in aqueous buffers. The organic solvent should not exceed 10% of the total react volume.
Is a hazy solution common after addition of the dissolved DSS solution to an aqueous buffer solution?

Yes. The resultant reaction mixture will become hazy frequently after the addition of the reagent solution.
How much DSS crosslinking reagent should I use?

We typically use a 10-fold molar excess of crosslinker to protein for solutions greater than 5 mg/mL. Use a 20- to 50-fold molar excess for more dilute protein solutions.
How is the DSS crosslinking reaction quenched?

The reaction is quenched by the addition of 1 M Tris or glycine, pH 7.5 to the reaction mixture to a final concentration of 20-50 mM.
Can I make a stock solution of the DSS crosslinker for long-term storage?

Stock solutions for long-term storage are NOT recommended because NHS esters are extremely vulnerable to hydrolysis. Even organic solvents are hygroscopic and tend to absorb water and promote hydrolysis. The crosslinker should be made immediately before each use.
What is the protocol for crosslinking IgG to Dynabeads from the Dynabeads Protein G Immunoprecipitation Kit (Cat. No. 10007D) using DSS (disuccinimidyl suberate)?

For crosslinking the antibody to Protein G Dynabeads, please refer to the crosslinking protocol with BS3 (bis(sulfosuccinimidyl)suberate) or Sulpha-DSS, which is an analogue of DSS. The only difference between these two molecules is that DSS is water insoluble and needs to be solubilized in DMSO before use. You can find the protocol at the following link.
Can you provide the shelf-life for DSS (disuccinimidyl suberate)?

DSS (disuccinimidyl suberate) 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.

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

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