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  6. Thermo Scientific™ Sulfo-SBED Biotin Label Transfer Reagent, No-Weigh™ Format

Thermo Scientific™ Sulfo-SBED Biotin Label Transfer Reagent, No-Weigh™ Format

Catalog No. PI33033
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10 mg
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Catalog No. Quantity
PI33033 10 mg
PIA39260 10 x 1 mg
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Catalog No. PI33033 Supplier Thermo Scientific™ Supplier No. 33033
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Description

Chemically tag purified proteins at primary amines to covalently crosslink and transfer biotinylation to protein interaction binding partners.

Thermo Scientific™ Pierce Sulfo-SBED Biotin Label Transfer Reagent is a multifunctional reagent for labeling a purified protein and then covalently transferring the attached biotin tag onto specific interactors of that protein.

Ready-to-use kit eliminates the need to prepare the necessary buffers or purchase the critical reagents necessary to perform a label transfer experiment. Key reagents, Sulfo-SBED and DTT, are supplied in unique, no-weigh, single-use packaging.

Highlights:

  • Sulfo-NHS ester reacts with primary amines at pH 7-9
  • Aryl azide conjugates randomly when activated with UV light
  • Cleavable disulfide bond allows biotin label transfer
  • Discover protein interactions without radiolabeling

Includes:

No-Weigh Sulfo-SBED Label Transfer Reagent, 8 × 1mg; BupH Phosphate Buffered Saline, 1 Pk. (makes 500mL); Label Transfer Buffer (20X), 200mL (makes 4L); Streptavidin-HRP Conjugate, 0.1mg; No-Weigh Dithiothreitol (DTT), 8 × 7.7mg; Slide-A-Lyzer MINI Dialysis Units plus Float, 10K MWCO, 10-100μL, 10 units/Pk.

TRUSTED_SUSTAINABILITY

Specifications

Format Powder
Quantity 10 mg
Product Type Crosslinker
Solubility Water
Content And Storage Upon receipt store at 4°C.
Labeling Method Chemical Labeling, Label Transfer
Reactive Moiety Sulfo-NHS Ester, Aryl Azide
Product Line Pierce

Frequently Asked Questions (FAQs)

What makes Sulfo-SBED Biotin Label Transfer Reagent useful for investigation of protein interactions?

Sulfo-SBED allows one to transfer a biotin tag from an amine-containing protein to a second protein (reacted with the phenyl azide). The amine-containing protein can be removed by reduction of the disulfide in the Sulfo-SBED spacer arm. This leaves just the second biotinylated protein for detection.
Should the Sulfo-SBED coupling reaction be protected from light?

During the initial coupling procedure, the solutions should be protected from light to avoid decomposition of the phenyl azide functional group.
Which reaction should take place first during Sulfo-SBED crosslinking?

The NHS-ester end of the crosslinker is subject to hydrolysis, therefore the amine-containing protein should be conjugated prior to the photoactivatable (aryl azide) reaction.
What is the appropriate wavelength for the conjugation of the photoactivatable group in Sulfo-SBED Biotin Label Transfer Reagent ?

UV light at wavelengths greater than 300 nm are appropriate for conjugation.
Is the Sulfo-SBED crosslinker soluble in organic solutions as well as aqueous solutions?

Yes, Sulfo-SBED is soluble in aqueous as well as organic solutions. It is most soluble in organics such as DMF and DMSO.
Can Sulfo-SBED Biotin Label Transfer Reagent be stored in a stock solution?

No, the crosslinker is moisture sensitive. The sulfo-NHS ester is easily hydrolyzed.
Can you provide the shelf-life for Sulfo-SBED Biotin Label Transfer Reagent?

Sulfo-SBED Biotin Label Transfer Reagent 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.
Can you provide the shelf-life for the Sulfo-SBED Label Transfer Reagent?

The Sulfo-SBED Biotin Label Transfer Reagent 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.
What crosslinkers do you recommend to study protein-protein interactions that are mass spectrometry - compatible?

We offer Sufo-SBED (Cat. Nos. 33033 and A39260).
What can I do if after label transfer of my sample with sulfo-SBED, I do not detect any signal in western blot?

  1. Determine the label incorporation of the bait protein before photoactivation with the prey protein, by western blot analysis.
  2. Check the UV light source to ensure good photoactivation (see Photolysis Information section of the Sulfo-SBED manual (https://tools.thermofisher.com/content/sfs/manuals/MAN0011509_SulfoSBED_BiotinLabel_Xsfer_WestBlotApp_UG.pdf) for guidelines).
  3. If the target (prey) protein concentration is too low, enrich the protein system if possible.
  4. Optimize the antibody concentrations used in western blot.
  5. It is possible that the protein interaction is blocking the epitope, which may occur with monoclonal antibodies and peptide-directed antibodies; in this case, use an antibody from a different source or probe with Streptavidin-HRP.
  6. Milk contains endogenous biotin and its use as a blocking buffer during the detection assay could mask the bands of the biotin-labeled interaction partner.
During label transfer using the sulfo-SBED Biotin Label Transfer Reagent, how is the spacer arm cleaved to separate the bait protein from the label?

In the case of sulfo-SBED biotin, there is a disulfide in the spacer arm of this tri-functional crosslinker that is cleaved with reducing agents such as DTT, beta-ME or TCEP. As the disulfide lies between the contact point on the bait protein and the complex of the photoreactive group and biotin, reduction separates the bait protein from the prey protein; which is now biotin labeled.
During label transfer using the sulfo-SBED Biotin Label Transfer Reagent, what groups are targeted on the bait and prey protein?

The NHS-ester of sulfo-SBED biotin will react with primary amines on the bait protein. The photoreactive aryl azide primarily targets nucleophiles, such as -NH2, -SH and -OH, but in their absence, it will react with any near neighbor molecule.
What are the label transfer reagents you offer?

Currently, we offer sulfo-SBED Biotin Label Transfer Reagent (Cat. Nos. 33033 and 33034) as well as the Sulfo-SBED Biotin Label Transfer Kit for Western Blot Application (Cat. No. 33073).

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

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