SPDP and LC-SPDP

Thermo Scientific Pierce SPDP is a short-chain crosslinker for amine-to-sulfhydryl conjugation via NHS-ester and pyridyldithiol reactive groups that form cleavable (reducible) disulfide bonds with cysteine sulfhydryls.

Thermo Scientific Pierce LC-SPDP is a long-chain crosslinker for amine-to-sulfhydryl conjugation via NHS-ester and pyridyldithiol reactive groups that form cleavable (reducible) disulfide bonds with cysteine sulfhydryls.

We offer three analogs of SPDP: the standard version (SPDP), a derivative with a longer spacer arm (LC-SPDP), and a sulfonated water-soluble variety (Sulfo-LC-SPDP). SPDP (N -Succinimidyl 3-(2-pyridyldithio)-propionate) and LC-SPDP (Succinimidyl 6-(3-[2-pyridyldithio]-propionamido)hexanoate) are heterobifunctional, thiol-cleavable and membrane permeable crosslinkers. They each contain an amine-reactive N-hydroxysuccinimide (NHS) ester that will react with lysine residues to form a stable amide bond. The other end of the spacer arm is terminated in the pyridyl disulfide group that will react with sulfhydryls to form a reversible disulfide bond.

Highlights:

• Releases a detectable by-product when reacted with a free sulfhydryl group; by measuring the release of pyridine-2-thione at 343 nm, the reaction can be easily followed
• Amine reactive Sulfo-NHS ester reacts rapidly with any primary amine-containing molecule
• Disulfide bond in the spacer arm is readily cleaved by 10-50 mM DTT or TCEP at pH 8.5
• Spacer arm is also cleaved with 5% β-mercaptoethanol in SDS-PAGE sample loading buffer at 100°C for 5 minutes
• Cleavable crosslinker allows separation of crosslinked products
• SPDP crosslinker is membrane permeable, so crosslinking can be carried out inside the cell

Applications:

• Reversible crosslinking of proteins
• Adding sulfhydryl groups to proteins by attachment to lysine residues
• Protein crosslinking to create antibody-enzyme and other bioconjugates via a two-step reaction
• Attaching haptens to carrier proteins for antibody production
• Preparing immunotoxin conjugates

Product Details:

Specifications:

We manufacture SPDP and LC-SPDP to the highest specifications to produce the most specific bioconjugates, ensure the integrity of your data and to provide you with the highest degree of consistency. Each lot of SPDP and LC-SPDP is tested to ensure it is >95% pure by quantitative NMR (the highest standard for crosslinker purity).

References:

1. Carlsson, J., et al. (1978) Protein thiolation and reversible protein-protein conjugation. N-succinimidyl-3-(2-pyridyldithio)propionate, a new heterobifunctional reagent. Biochem. J. 173 , 723-737.
2. Cumber, A.J., et al. (1985) Preparation of antibody-toxin conjugates. Meth. Enzymol . , New York : Academic Press, 112, 207-225.
3. Engstler, M., et al. (2004) J. Cell Sci.117, 1105-1115.
4. Itoh, Y., et al. (2001) Proc. Natl. Acad. Sci. USA98, 4883-4887.
5. Wang, D., et al . (1997) Bioconjug. Chem. 8 , 878-884.

Related Products:

Sulfo-LC-SPDP
PEG4-SPDP
PEG12-SPDP