Name: DSP Crosslinker
Brand: BOC Sciences
Price: 680 USD
Availability: MadeToOrder

Synonyms

Disuccinimido dithiobispropionate; Di(N-succinimidyl) 3,3'-dithiodipropionate; Dithiobis(succinimidylpropionate); DTSP; Bis(2,5-dioxopyrrolidin-1-yl) 3,3'-disulfanediyldipropanoate

IUPAC Name

(2,5-dioxopyrrolidin-1-yl) 3-[[3-(2,5-dioxopyrrolidin-1-yl)oxy-3-oxopropyl]disulfanyl]propanoate

Canonical SMILES

C1CC(=O)N(C1=O)OC(=O)CCSSCCC(=O)ON2C(=O)CCC2=O

InChI

InChI=1S/C14H16N2O8S2/c17-9-1-2-10(18)15(9)23-13(21)5-7-25-26-8-6-14(22)24-16-11(19)3-4-12(16)20/h1-8H2

InChIKey

FXYPGCIGRDZWNR-UHFFFAOYSA-N

Density

1.570±0.10 g/cm<sup>3</sup> (Predicted)

Solubility

10 mm in DMSO

Melting Point

128-133 °C

Flash Point

292.6±32.9 °C

Index Of Refraction

1.625

PSA

177.96000

Vapor Pressure

0.0±1.5 mmHg at 25°C

Appearance

Off-White Solid

Shipping

Room temperature

Storage

Store at 2-8 °C

Pictograms

Irritant

Signal Word

Warning

Boiling Point

560.1±60.0 °C (Predicted)

In Vivo

Crosslinked DNA polyplexes showed an increased stability against exchange reaction by salt or heparin. Disulfide bond containing DSP-linked polyplexes were susceptible to reducing conditions. These polyplexes displayed the highest gene expression levels in vitro and in vivo (upon intratumoral application in mice), and these were significantly elevated and prolonged over standard or DSS-stabilized HD O formulations. DSP-stabilized HD O polyplexes with or without Tf coating were well-tolerated after intravenous application. High gene expression levels were found in tumor tissue, with negligible gene expression in any other organ.

Biological Activity

DSP Crosslinker is a cleavable ADC linker, used in the synthesis of antibody-drug conjugates (ADCs)[1] . In Vitro: ADCs are comprised of an antibody to which is attached an ADC cytotoxin through an ADC linker

DSP Crosslinker, a chemical reagent facilitating covalent linkages between molecules, is an indispensable tool in a multitude of scientific pursuits. Here are four key applications of DSP Crosslinker:

Protein-Protein Interaction Studies: Widely utilized in the exploration of protein-protein interactions, DSP Crosslinker serves to forge stable complexes between interacting proteins. By covalently linking these proteins, researchers can capture fleeting interactions, subsequently subjecting the complex to analysis through methodologies like mass spectrometry or gel electrophoresis. This approach aids in the identification of interaction partners and enhances comprehension of the intricate functional networks operating within cells.

Immunoprecipitation Assays: Within immunoprecipitation assays, DSP Crosslinker plays a crucial role in fortifying antibody-antigen complexes. By crosslinking the antibody to the antigen, the resulting complex gains heightened resistance to dissociation during washing stages, elevating specificity and yield in the assay. This augmentation strengthens the reliability of immunoprecipitation as a potent investigative tool for scrutinizing protein function and localization.

Bioconjugation: Encompassing bioconjugation endeavors, DSP Crosslinker facilitates the attachment of biomolecules to diverse substrates, such as beads or surfaces. This process enables the fabrication of functionalized surfaces for applications spanning biosensors, diagnostic tools, and affinity purification systems. The capability to covalently link biomolecules ensures stability and specificity in these contexts, fostering advancements in biotechnological applications.

Structural Biology: Integral to the landscape of structural biology, DSP Crosslinker contributes to the crosslinking of proteins in their native conformations, facilitating structural studies utilizing techniques like X-ray crystallography or cryo-EM. By stabilizing the native protein complexes, researchers can glean more precise structural insights, a critical facet in unraveling protein functionality and devising therapeutic interventions.

1. DSP-crosslinking and Immunoprecipitation to Isolate Weak Protein Complex

Kotaro Akaki, Takashi Mino, Osamu Takeuchi Bio Protoc. 2022 Aug 5;12(15):e4478. doi: 10.21769/BioProtoc.4478.

Detecting protein-protein interactions (PPIs) is one of the most used approaches to reveal the molecular regulation of protein of interests (POIs). Immunoprecipitation of POIs followed by mass spectrometry or western blot analysis enables us to detect co-precipitated POI-binding proteins. However, some binding proteins are lost during cell lysis or immunoprecipitation if the protein binding affinity is weak. Crosslinking POI and its binding proteins stabilizes the PPI and increases the chance of detecting the interacting proteins. Here, we introduce the method of DSP (dithiobis(succinimidyl propionate))-mediated crosslinking, followed by tandem immunoprecipitation (FLAG and HA tags). The eluted proteins interacting with POI can be analyzed by mass spectrometry or western blotting. This method has the potential to be applied to various cytoplasmic proteins. Graphical abstract.

2. An improved CUT&RUN method for regulation network reconstruction of low abundance transcription factor

Huiru Bai, Meizhen Lin, Yuan Meng, Huiyuan Bai, Shang Cai Cell Signal. 2022 Aug;96:110361. doi: 10.1016/j.cellsig.2022.110361. Epub 2022 May 25.

By improving the previous method of CUT&RUN, we developed D-CUT&RUN (DSP fixed CUT&RUN) for under-expressed transcription factor. High-quality data could be obtained for low expressed transcription factors using chemical crosslinkers (DSP) and reducing agent (DTT). We applied our D-CUT&RUN to detection of Bcl11b and Mycn binding sites in mammary epithelial progenitor cells. Pathway enrichment analysis results of Bcl11b target genes showed that Bcl11b was a regulatory factor involved in breast cancer and it could negatively regulate Wnt signaling pathway. Furthermore, the role of Bcl11b in breast cancer was mediated by catabolic process and stress-related pathway. Our research suggested that D-CUT&RUN could be used for low abundance transcription factor binding sites detection and Bcl11b could be a target for breast cancer treatment in the future.

3. Structural and Functional Insights into CP2c Transcription Factor Complexes

Seung Han Son, Min Young Kim, Eunbi Jo, Vladimir N Uversky, Chul Geun Kim Int J Mol Sci. 2022 Jun 7;23(12):6369. doi: 10.3390/ijms23126369.

CP2c, also known as TFCP2, α-CP2, LSF, and LBP-1c, is a prototypic member of the transcription factor (TF) CP2 subfamily involved in diverse ubiquitous and tissue/stage-specific cellular processes and in human malignancies including cancer. Despite its importance, many fundamental regulatory mechanisms of CP2c are still unclear. Here, we uncover unprecedented structural and functional aspects of CP2c using DSP crosslinking and Western blot in addition to conventional methods. We found that a monomeric form of a CP2c homotetramer (tCP2c; [C4]) binds to the known CP2c-binding DNA motif (CNRG-N(5~6)-CNRG), whereas a dimeric form of a CP2c, CP2b, and PIAS1 heterohexamer ([C2B2P2]2) binds to the three consecutive CP2c half-sites or two staggered CP2c binding motifs, where the [C4] exerts a pioneering function for recruiting the [C2B2P2]2 to the target. All CP2c exists as a [C4], or as a [C2B2P2]2 or [C2B2P2]4 in the nucleus. Importantly, one additional cytosolic heterotetrameric CP2c and CP2a complex, ([C2A2]), exerts some homeostatic regulation of the nuclear complexes. These data indicate that these findings are essential for the transcriptional regulation of CP2c in cells within relevant timescales, providing clues not only for the transcriptional regulation mechanism by CP2c but also for future therapeutics targeting CP2c function.

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