Dithiobis succinimidyl propionate dsp

Manufactured by Merck Group

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Dithiobis(succinimidyl propionate) (DSP) is a crosslinking reagent used in biochemistry and molecular biology applications. It is a homobifunctional N-hydroxysuccinimide (NHS) ester that can form covalent bonds between primary amino groups. DSP is often used to conjugate proteins, peptides, or other molecules in various experiments and assays.

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Lab products found in correlation

Hydroxylamine, by Merck Group (1 mentions) Tween 20, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) L methionine, by Thermo Fisher Scientific (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Ab213831, by Abcam (1 mentions) N hydroxysulfosuccinimide sulfo nhs, by Thermo Fisher Scientific (1 mentions) Anti cd63 ab59479, by Abcam (1 mentions) Tfmba, by Merck Group (1 mentions) Ascorbic acid, by MP Biomedicals (1 mentions) Hydrogen tetrachloroaurate 3 trihydrate, by Merck Group (1 mentions) 11 mercaptoundecanoic acid mua, by Merck Group (1 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride edc, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Propionate (76 citations) Dithiobis-[Succinimidyl Propionate]) (3 citations)

6 protocols using dithiobis succinimidyl propionate dsp

Antibody-Based Detection of E. coli and MS2 Bacteriophage

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Optical qPCR tubes (8× strip, catalog
no. 401428) and separate caps (8× strip, catalog no. 401425)
were obtained from Agilent Technologies (Santa Clara, CA). Rabbit
polyclonal anti-E. coli antibodies were obtained
from Fitzgerald (Acton, MA). Rabbit polyclonal anti-MS2 antibodies
were obtained from Tetracore (Rockville, MD). E. coli (strain MB1457), and bacteriophage MS2 (strain 15597-B1) were obtained
from the American Type Culture Collection (Manassas, VA). Phosphate
buffered saline (PBS) tablets (10 mM Phosphate, 2.7 mM potassium chloride,
140 mM sodium chloride, pH 7.4) were purchased from Bioline (Taunton,
MA). Anonymized human serum was obtained from the Gulf Coast Regional
Blood Center (Houston, TX). Fluorescein isothiocyanate (FITC) was
obtained from Pierce (Rockford, IL). 1-Ethyl-3-(3-(dimethylamino)propyl)
carbodiimide hydrochloride (EDC), N-hydroxysuccinimide
(NHS), bovine serum albumin (BSA), Tween-20, hydroxylamine, 6-mercapto-1-hexanol,
dimethyl sulfoxide (DMSO), anhydrous chromium trioxide, 96.7% sulfuric
acid, dithiobis(succinimidyl propionate) (DSP), and sodium cyanoborohydride
were purchased from Sigma-Aldrich (St. Louis, MO).

Garvey G., Shakarisaz D., Ruiz-Ruiz F., Hagström A.E., Raja B., Pascente C., Kar A., Kourentzi K., Rito-Palomares M., Ruchhoeft P, & Willson R.C. (2014). Microretroreflector-Sedimentation Immunoassays for Pathogen Detection. Analytical Chemistry, 86(18), 9029-9035.

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Photoactivatable Amino Acid Labeling and Chemical Crosslinking

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Photoactivatable L-leucine and L-methionine were purchased from Thermofisher. These analogs were substituted at 2 mM final concentration to replace natural amino acids. After proteoliposome purification, the samples aliquoted in microtubes were subjected to ultra violet (UV) light at 302 nm for one minute. The reaction was quenched by incubation with 1.5 M Tris-HCl, pH 7.5, at room temperature for 15 min. Analysis of the samples was performed by SDS-PAGE.
Dithiobis(succinimidyl propionate) (DSP) was purchased from Sigma-Aldrich. Proteoliposomes were incubated for 30 minutes at room temperature with 5 mM of DSP. The reaction was quenched by incubation with 1.5 M Tris-HCl, pH 7.5, at room temperature for 15 min. SDS-PAGE analysis of cross-linked samples was performed under non-reducing conditions.

Soranzo T., Cortès S., Gilde F., Kreir M., Picart C, & Lenormand J.L. (2015). Functional Characterization of p7 Viroporin from Hepatitis C Virus Produced in a Cell-Free Expression System. Protein expression and purification, 118, 83-91.

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Electrochemical Sensor Functionalization for cTnI Detection

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Dithiobis succinimidyl propionate (DSP) (Sigma-Aldrich, St. Louis, MO, USA) was dissolved in dimethyl sulfoxide (DMSO) (Sigma-Aldrich, St. Louis, MO, USA) to formulate a 10 mM mixture. Thirty microliters of the DSP-DMSO mixture was added to the Mo electrochemical sensor to allow functionalization of this thiol-based linker molecule on Mo surface and incubated for four hours. Phosphate-buffered saline (PBS) buffer (0.15 M) was added to the sensor to prepare the surface prior to addition of the antibody. Monoclonal anti-cTnI antibody stock solution was diluted to 1 μg/mL in PBS buffer and then immobilized on the DSP functionalized sensor surface and incubated for 15 min. The concentration of antibody to be used was determined through an antibody saturation study. The antibody saturation study experiment was conducted with varying antibody concentrations from 100 fg/mL to 1 μg/mL and the change in impedance with respect to a blank PBS sample was studied for the various antibody concentrations. The noise estimation after-antibody conjugation was studied by analyzing the impedance of the sensor for multiple PBS washes following the antibody conjugation. EIS measurements were taken after each assay step with Gamry Reference 3000 potentiostat (Gamry Instruments, Warminster, PA, USA) to validate the binding.

Kamakoti V., Panneer Selvam A., Radha Shanmugam N., Muthukumar S, & Prasad S. (2016). Flexible Molybdenum Electrodes towards Designing Affinity Based Protein Biosensors. Biosensors, 6(3), 36.

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In vivo Cross-linking of Laz and AniA in N. gonorrhoeae

Cited 1 time

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To evaluate interactions between Laz and AniA, an in vivo cross-linking protocol (Gray et al., 2011 (link)) was adapted for N. gonorrhoeae. Non-piliated bacteria were collected from GCB and suspended to a final OD₆₀₀ of 1.5 in 4 mL sterile PBS. Suspensions were centrifuged, supernatants were decanted, and pellets were suspended in 500 μL sterile PBS. Dithiobis(succinimidyl propionate) (DSP; Sigma) was hydrated to 25 mM in dimethylsulfoxide and added to 100 μL of each suspension to 0, 0.1, 0.25, or 0.4 mM. Cross-linking reactions were incubated at 37°C and 5% CO₂ for 30 min and quenched by the addition of 50 mM Tris-HCl pH 8.0 and incubation at room temperature for 10 min. Cells were lysed by boiling in sample buffer without reducing agent to avoid cleaving DSP-linked complexes. SDS-PAGE separation was performed at 4°C and 150 V followed by immunoblotting.

Baarda B.I., Zielke R.A., Jerse A.E, & Sikora A.E. (2018). Lipid-Modified Azurin of Neisseria gonorrhoeae Is Not Surface Exposed and Does Not Interact With the Nitrite Reductase AniA. Frontiers in Microbiology, 9, 2915.

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Peptide Binding Assay Protocols

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Dithiobis(succinimidyl propionate) (DSP) was purchased from Sigma-Aldrich (St. Louis, MO). Peptides utilized in a surface binding assay study were CLIP (Ii_97–120) (₉₇LPKPPKPVSKMRMATPLLMQALPM₁₂₀) and tetanus toxin_947–967 (TT_947–967) (₉₄₇FNNFTVSFWLRVPKVSASHLE₉₆₇). Peptides used in the competitive binding assay were HIV ENV_31–45 (₃₁ TEKLWVTVYYGVPVW₄₅), MAGE-A3_243–258 (₂₄₃KKLLTQHFVQENYLEY₂₅₈), N-extended CLIP (₉₂NLRMKLPKPPKPVSKMRMATPLLMQALPM₁₂₀), C-extended CLIP (₉₇LPKPPKPVSKMRMATPLLMQALPMGALPQ₁₂₅), and CLIP (as above). All peptides were purchased from GenScript (Piscataway, NJ).

Anczurowski M., Yamashita Y., Nakatsugawa M., Ochi T., Kagoya Y., Guo T., Wang C.H., Rahman M.A., Saso K., Butler M.O, & Hirano N. (2018). Mechanisms underlying the lack of endogenous processing and CLIP-mediated binding of the invariant chain by HLA-DP84Gly. Scientific Reports, 8, 4804.

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Characterization of Extracellular Vesicles

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Hydrogen tetrachloroaurate(III) trihydrate (HAuCl₄·3H₂O), silver nitrate (AgNO₃), MBA, DTNB, TFMBA, MMC, dithiobis (succinimidyl propionate) (DSP), and 11‐mercaptoundecanoic acid (MUA) were obtained from Sigma–Aldrich. Ascorbic acid (AA) was purchased from MP Biomedicals, Inc. 1‐Ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N‐hydroxysulfosuccinimide (Sulfo‐NHS) were bought from Thermo Fisher Scientific. Anti‐CD63 (ab59479) was ordered from Abcam. THSB2 (MAB16351) was purchased from R&D Systems, and VCAN (NBP1‐85432) and TNC (NOVNB11068136) were obtained from Novus Biologicals. THBS2, VCAN, and TNC ELISA kits were bought from Invitrogen (EH452RB), R&D Systems (NBP2‐75354), and Abcam (ab213831), respectively.

Li J., Sina A.A., Antaw F., Fielding D., Möller A., Lobb R., Wuethrich A, & Trau M. (2022). Digital Decoding of Single Extracellular Vesicle Phenotype Differentiates Early Malignant and Benign Lung Lesions. Advanced Science, 10(1), 2204207.

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