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DSS (disuccinimidyl suberate), 1 g - FAQs

DSS (disuccinimidyl suberate), 1 g - FAQs

查看更多产品信息 DSS (disuccinimidyl suberate), No-Weigh™ Format - FAQs (21655, 21555, A39267)

12 个常见问题解答

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).

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

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

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.

Find additional tips, troubleshooting help, and resources within our Protein Immunoprecipitation (IP), Co-Immunoprecipitation (Co-IP), and Pulldown Support Center

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.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.