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N hydroxysulfosuccinimide

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N-hydroxysulfosuccinimide is a water-soluble compound commonly used as an ester-forming reagent in biochemical applications. It serves as an activating agent in the conjugation of various molecules, such as proteins, peptides, and other biomolecules.

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Lab products found in correlation

1 ethyl 3 3 dimethylaminopropyl carbodiimide, by Thermo Fisher Scientific (4 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide edc, by Thermo Fisher Scientific (3 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride edc, by Thermo Fisher Scientific (2 mentions) 1 ethyl 3 3 dimethylaminopropyl carbodiimide hydrochloride, by Thermo Fisher Scientific (2 mentions) Sodium azide, by Avantor (2 mentions) Tween 20, by Avantor (2 mentions) Bovine serum albumin (bsa), by Boston BioProducts (2 mentions) Nah2po4, by Avantor (2 mentions) T20 cell counter, by Bio-Rad (2 mentions) Phosphate buffered saline (pbs), by Avantor (2 mentions) Aptes, by Merck Group (2 mentions) Carboxyl polystyrene beads, by Spherotech (1 mentions) 11 mercaptoundecanoic acid, by Merck Group (1 mentions) Gold 3 chloride hydrate haucl4, by Merck Group (1 mentions) Hexadecyltrimethylammonium bromide ctab, by Tokyo Chemical Industry (1 mentions) Human il 10 uncoated elisa kit, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Anhydrous ethanol, by Merck Group (1 mentions) Superblock t20 blocking buffer, by Thermo Fisher Scientific (1 mentions) Hydrochloric acid (hcl), by Merck Group (1 mentions)

Close spelling variants of this product

N-hydroxysulfosuccinimide (Sulfo-NHS) (46 citations) N-hydroxysulfosuccinimide sodium salt (Sulfo-NHS) (9 citations) Sulfo-N-Hydroxysulfosuccinimide (8 citations) N-hydroxysulfosuccinimide (NHS) (3 citations) N-hydroxysulfosuccinimide sodium salt (NHS, (3 citations)

20 protocols using n hydroxysulfosuccinimide

1

Biomimetic Microfluidic System for Drug Delivery

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Human serum albumin (HSA), oleic acid (OA), and DOX were purchased from Sigma-Aldrich (St. Louise, MO, USA). PTX was obtained from Daewoong Pharmaceutical Co., Ltd. (Seoul, Republic of Korea). 1-Ethyl-3-(3-dimethylamino-propyl) and N-hydroxysulfosuccinimide were purchased from Thermo Fisher Scientific Korea (Seoul, Republic of Korea). In contrast, biomimetic microfluidic system components were purchased from BioSpero Co., Ltd. (Jeju, Republic of Korea). Deionized water was processed using ultrapure water systems (Mirae ST Co., Ltd., Anyang, Republic of Korea). Other high-performance liquid chromatography (HPLC)-grade organic solvents were purchased from Samchundang Chemicals (Seoul, Republic of Korea).
Kim H., Kim E.J., Ngo H.V., Nguyen H.D., Park C., Choi K.H., Park J.B, & Lee B.J. (2023). Cellular Efficacy of Fattigated Nanoparticles and Real-Time ROS Occurrence Using Microfluidic Hepatocarcinoma Chip System: Effect of Anticancer Drug Solubility and Shear Stress. Pharmaceuticals, 16(9), 1330.
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2

Antibody-Conjugated Quantum Dots Synthesis

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The anti-CD11b antibody (H5A4) supernatant, developed by J.T. August and J.E.K. Hildreth of Johns Hopkins University, was obtained from the Developmental Studies Hybridoma Bank, created by the NICHD of the NIH and maintained at the University of Iowa, Department of Biology, Iowa City, IA 52242. Anti-CD11b-conjugated AIS/Zn QDs, referred to as CD11b-QD, were prepared by covalently conjugating anti-CD11b antibody (H5A4) to AIS/Zn QDs using EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride; Thermo Fisher Scientific) / NHS (N-hydroxysulfosuccinimide; Thermo Fisher Scientific) chemistry. Specifically, as-synthesized AIS/Zn QD solutions were sterilized by passage through a 0.2 μm PES filter. The QDs in Tris-HCl buffer (pH 9) were then re-suspended in 1X phosphate-buffered saline (PBS; pH 7.4), followed by three washes in PBS using 30-kDa Amicon centrifugal filters (4500 g, 5 min). 500 μL of a 20 mg/mL EDC solution and 500 μL of a 20 mg/ml NHS solution were added to 5 ml of the sterilized AIS/Zn QDs in PBS solution, and the solution was incubated for 30 min at room temperature. Conjugation of the anti-CD11b antibody to AIS/Zn QDs was initiated by the addition of 50 μl of anti-CD11b antibody, followed by incubation for 30 min at room temperature.
Ozdemir N.K., Cline J.P., Sakizadeh J., Collins S.M., Brown A.C., McIntosh S., Kiely C.J, & Snyder M.A. (2022). Sequential, Low-Temperature Aqueous Synthesis of Ag-In-S/Zn Quantum Dots via Staged Cation Exchange under Biomineralization Conditions. Journal of materials chemistry. B, 10(24), 4529-4545.
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3

Functionalization of Polystyrene Microspheres with DNA

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To functionalize polystyrene microspheres with DNA, we mixed either single-stranded or double-stranded oligonucleotides containing a 5’ amino-modifier (Table s2) with 2.1 μm carboxyl-polystyrene beads (Spherotech). Coupling of the oligonucleotide to the bead surface was achieved by chemical crosslinking with 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride (EDC; ThermoFisher) in the presence of N-hydroxysulfosuccinimide (Sulfo-NHS) in 100 mM MES-Na (pH 6) buffer. The reaction was quenched by addition of TE8 buffer (20 mM Tris-HCl, 1 mM EDTA, pH 8). Unreacted material was removed by washing the beads three times with TE8. Beads modified with single-stranded DNA (“ssDNA-beads”) were used to generate long DNA handles. Beads modified with double-stranded DNA (“dsDNA-beads”) were used to immobilize proteins and RNCs. In this case, the double-stranded oligonucleotides contained a 4-nucleotide single-stranded overhang with a sequence reverse-complementary to the overhang of the dsOligo-CoA DNA.
Liu K., Maciuba K, & Kaiser C.M. (2019). The ribosome cooperates with a chaperone to guide multi-domain protein folding. Molecular cell, 74(2), 310-319.e7.
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4

Gold Nanoparticle-Based IL-10 ELISA

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The human IL-10 uncoated ELISA kit including IL-10 protein, IL-10 capture antibody, and IL-10 detection antibody was purchased from Invitrogen. Gold (III) chloride hydrate (HAuCl4), sodium borohydride (NaBH4), L-ascorbic acid, 11-mercaptoundecanoic acid (MUA), and bovine serum albumin (BSA) were purchased from Sigma-Aldrich. 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysulfosuccinimide (NHSS) were purchased from Thermo Fisher Scientific Inc. Hexadecyltrimethylammonium bromide (CTAB) was purchased from Tokyo Chemical Industry Co., Ltd. Phosphate-buffered saline (PBS) for biofunctionalization of AuNC was purchased from Biosesang.
Baek S.H., Song H.W., Lee S., Kim J.E., Kim Y.H., Wi J.S., Ok J.G., Park J.S., Hong S., Kwak M.K., Lee H.J, & Nam S.W. (2020). Gold Nanoparticle-Enhanced and Roll-to-Roll Nanoimprinted LSPR Platform for Detecting Interleukin-10. Frontiers in Chemistry, 8, 285.
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5

Materials for Bioanalytical Assay Examples

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Example 1

The following example describes materials used in Examples 2-10. Optical fiber bundles were purchased from Schott North America (Southbridge, Mass.). Non-reinforced gloss silicone sheeting was obtained from Specialty Manufacturing (Saginaw, Mich.). Hydrochloric acid, anhydrous ethanol, and molecular biology grade Tween-20 were purchased from Sigma-Aldrich (Saint Louis, Mo.). 2.7-μm-diameter carboxyl-terminated magnetic beads were purchased from Varian, Inc. (Lake Forest, Calif.). 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC), N-hydroxysulfosuccinimide (NHS), and SuperBlock® T-20 Blocking Buffer were purchased from Thermo Scientific (Rockford, Ill.). Streptavidin-β-galactosidase (SβG) was purchased from Invitrogen, Sigma-Aldrich, or conjugated in house using standard protocols. Resorufin-3-D-galactopyranoside (RGP) was purchased from Invitrogen (Carlsbad, Calif.). The fiber polisher and polishing consumables were purchased from Allied High Tech Products (Rancho Dominguez, Calif.). Monoclonal capture antibody to PSA, monoclonal detection antibody to PSA, and purified PSA were purchased from BiosPacific. The Chromalink™ biotinylation reagent was purchased from Solulink, Inc (San Diego, Calif.). Purified DNA was purchased from Integrated DNA Technologies.

US10989713B2. Methods and systems for extending dynamic range in assays for the detection of molecules or particles (2021-04-27). Quanterix Corporation [US]. Inventors: David M. Rissin [US], David Fournier [US], David C. Duffy [US].
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6

SARS-CoV-2 Protein Coupling to Microspheres

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SARS-CoV-2 proteins were coupled to MagPlex Microspheres of spectrally distinct regions (Luminex; Austin, TX). Coupling was carried out at room temperature following standard carbodiimide coupling procedures. Microspheres were washed once with deionized water and incubated for 20 min in the dark in an end-over-end rotator suspension of 0.1 M NaH2PO4 (VWR International; Radnor, PA), 5 mg/ml 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (Thermo Fisher Scientific; Waltham, MA), and 5 mg/ml N-hydroxysulfosuccinimide; Thermo Fisher Scientific). Activated microspheres were washed twice in 0.05 M MES (Boston Bioproducts, Ashland, MA) and incubated for 2 h in the dark in an end-over-end rotator, 500-μl suspension of 0.05 M MES containing 300 nmol/l Ag and 1 × 106 microspheres (0.15 nmol Ag/106 microspheres). Coupled microspheres were washed four times in a blocking buffer consisting of 1% BSA (Boston Bioproducts), 1× PBS (VWR International), 0.05% sodium azide (VWR International), and 0.02% Tween 20 (VWR International). Coupled microspheres were then stored at a concentration of 106 spheres/ml at 4°C in the same blocking buffer. Concentrations of spheres were confirmed by counting on a Bio-Rad T20 Cell Counter.
Haddad N.S., Nguyen D.C., Kuruvilla M.E., Morrison-Porter A., Anam F., Cashman K.S., Ramonell R.P., Kyu S., Saini A.S., Cabrera-Mora M., Derrico A., Alter D., Roback J.D., Horwath M., O’Keefe J.B., Wu H.M., Wong A.K., Dretler A.W., Gripaldo R., Lane A.N., Wu H., Chu H.Y., Lee S., Hernandez M., Engineer V., Varghese J., Patel R., Jalal A., French V., Guysenov I., Lane C.E., Mengistsu T., Normile K.E., Mnzava O., Le S., Sanz I., Daiss J.L, & Lee F.E. (2021). One-Stop Serum Assay Identifies COVID-19 Disease Severity and Vaccination Responses. ImmunoHorizons, 5(5), 322-335.
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7

Multiplex SARS-CoV-2 and Influenza Antibody Assay

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Microspheres (Luminex) were coupled by 1-ethyl-3-[3-dimethylaminopropyl] carbodiimide hydrochloride and N-hydroxysulfosuccinimide (ThermoScientific) to RBD (BEI NR-52309), spike (BEI NR-52308), Delta spike (BEI NR-55614), Omicron spike (BEI NR-56447), and a mixture of influenza antigens (BEI NR-20083, NR-51702, NR-12148). Antigen-coupled Microspheres were incubated with sera. Bead-bound antigen-specific antibodies were detected using phycoerythrin (PE)-coupled anti-IgG/anti-IgA1/anti-IgM/anti-IgG1/anti-IgG2/anti-IgG3/anti-IgG4 (1 µg/mL; Southern Biotech). For FcR binding, FcγRIIIa/CD16a, FcγRIIa/CD32a, FcγRIIb/CD32b, and FcRN (R&D Systems) labeled with PE (Abcam) were used (1 µg/mL). PE signals were measured on a MAGPIX (Luminex). Replicate samples over 3 serial dilutions were tested in 2 independent experiments. Representative data were chosen by the highest signal-to-noise ratio [8 (link), 33 (link)].
Adhikari E.H., Lu P., Kang Y.J., McDonald A.R., Pruszynski J.E., Bates T.A., McBride S.K., Trank-Greene M., Tafesse F.G, & Lu L.L. (2023). Diverging Maternal and Cord Antibody Functions From SARS-CoV-2 Infection and Vaccination in Pregnancy. The Journal of Infectious Diseases, 229(2), 462-472.
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8

Antigen Immobilization on Magnetic Beads

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The viral peptides and proteins were immobilised on the surface of colour‐coded magnetic beads (MagPlex, Luminex Corp., Austin, TX, USA) as previously described.37 Briefly, each antigen was diluted to a final concentration of 80 µg mL−1 in 100 mm 2‐(N‐morpholino) ethanesulfonic acid buffer, pH 4.5 (Sigma‐Aldrich) and coupled to one bead identity (colour code). The carboxylated surface of 1 × 106 colour‐coded magnetic beads per bead identity was activated by using 100 µL phosphate buffer complemented with 0.5 mg 1‐(3‐dimethylaminopropyl)‐3‐ethylcarbodiimide hydrochloride (ProteoChem, Inc., Hurricane, UT, USA) and 0.5 mg N‐hydroxysulfosuccinimide (Thermo Fisher Scientific). Activated beads were incubated for 2 h with the diluted antigen, followed by overnight incubation in blocking buffer (Blocking Reagent for ELISA, Roche, supplemented with 0.1% (v/v) ProClin, Sigma‐Aldrich). The bead identities were then pooled to form the antigen‐bead array. Besides the viral antigens, two bead identities were coupled with anti‐human IgG (309‐005‐082, Jackson Immunoresearch, West Grove, PA, USA) and the EBNA1 protein (ab138345, Abcam, Cambridge, UK) from the common Epstein–Barr virus and used as sample loading controls.
Hober S., Hellström C., Olofsson J., Andersson E., Bergström S., Jernbom Falk A., Bayati S., Mravinacova S., Sjöberg R., Yousef J., Skoglund L., Kanje S., Berling A., Svensson A., Jensen G., Enstedt H., Afshari D., Xu L.L., Zwahlen M., von Feilitzen K., Hanke L., Murrell B., McInerney G., Karlsson Hedestam G.B., Lendel C., Roth R.G., Skoog I., Svenungsson E., Olsson T., Fogdell‐Hahn A., Lindroth Y., Lundgren M., Maleki K.T., Lagerqvist N., Klingström J., Da Silva Rodrigues R., Muschiol S., Bogdanovic G., Arroyo Mühr L.S., Eklund C., Lagheden C., Dillner J., Sivertsson Å., Havervall S., Thålin C., Tegel H., Pin E., Månberg A., Hedhammar M, & Nilsson P. (2021). Systematic evaluation of SARS‐CoV‐2 antigens enables a highly specific and sensitive multiplex serological COVID‐19 assay. Clinical & Translational Immunology, 10(7), e1312.
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9

Covalent Attachment of Complement Factor H to MSCs

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Primary MSCs were painted with purified human CFH (Complement technology Inc, Tyler, TX) after 1-ethyl-3-(-3-dimethylaminopropyl) carbodiimide hydrochloride (EDC)-mediated activation of carboxyl groups. In brief, 2 mM EDC and 5 mM N-hydroxysulfosuccinimide (ThermoFisher, Waltham, MA) were added to CFH solution at the indicated concentration and allowed to react for 15 min at room temperature to create activated carboxyl groups, then 20 mM β-mercaptoethanol was added to quench the excess EDC and the activated CFH was separated using a desalting column (ThermoFisher, Waltham, MA) equilibrated with PBS. MSCs (2×104) were then incubated with different concentrations of activated CFH for 30 min at 37°C in 100 μl PBS, then the cells were washed with PBS and used for experiments.
Li Y., Qiu W., Zhang L., Fung J, & Lin F. (2016). Painting factor H onto mesenchymal stem cells protects the cells from complement- and neutrophil-mediated damage. Biomaterials, 102, 209-219.
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10

SARS-CoV-2 Protein Coupling to Microspheres

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SARS-CoV-2 proteins were coupled to MagPlex Microspheres of spectrally distinct regions (Luminex; Austin, TX). Coupling was carried out at room temperature following standard carbodiimide coupling procedures. Microspheres were washed once with deionized water and incubated for 20 min in the dark in an end-over-end rotator suspension of 0.1 M NaH2PO4 (VWR International; Radnor, PA), 5 mg/ml 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (Thermo Fisher Scientific; Waltham, MA), and 5 mg/ml N-hydroxysulfosuccinimide; Thermo Fisher Scientific). Activated microspheres were washed twice in 0.05 M MES (Boston Bioproducts, Ashland, MA) and incubated for 2 h in the dark in an end-over-end rotator, 500-μl suspension of 0.05 M MES containing 300 nmol/l Ag and 1 × 106 microspheres (0.15 nmol Ag/106 microspheres). Coupled microspheres were washed four times in a blocking buffer consisting of 1% BSA (Boston Bioproducts), 1× PBS (VWR International), 0.05% sodium azide (VWR International), and 0.02% Tween 20 (VWR International). Coupled microspheres were then stored at a concentration of 106 spheres/ml at 4°C in the same blocking buffer. Concentrations of spheres were confirmed by counting on a Bio-Rad T20 Cell Counter.
Haddad N.S., Nguyen D.C., Kuruvilla M.E., Morrison-Porter A., Anam F., Cashman K.S., Ramonell R.P., Kyu S., Saini A.S., Cabrera-Mora M., Derrico A., Alter D., Roback J.D., Horwath M., O’Keefe J.B., Wu H.M., Wong A.K., Dretler A.W., Gripaldo R., Lane A.N., Wu H., Chu H.Y., Lee S., Hernandez M., Engineer V., Varghese J., Patel R., Jalal A., French V., Guysenov I., Lane C.E., Mengistsu T., Normile K.E., Mnzava O., Le S., Sanz I., Daiss J.L, & Lee F.E. (2021). One-Stop Serum Assay Identifies COVID-19 Disease Severity and Vaccination Responses. ImmunoHorizons, 5(5), 322-335.
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