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Nhs sulfo lc lc kit

Manufactured by Thermo Fisher Scientific
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The NHS-Sulfo-LC-LC kit is a crosslinking reagent used for protein modification and conjugation. It contains a water-soluble, cleavable linker with NHS ester groups on both ends, allowing for the covalent attachment of biomolecules.

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Lab products found in correlation

Zeba spin desalting column, by Thermo Fisher Scientific (6 mentions) Hcov hku1 spike, by Sino Biological (2 mentions) Hcov oc43 spike oc, by Sino Biological (2 mentions) Influenza haemagglutinin ha, by Sino Biological (2 mentions) Fluorescent neutravidin beads, by Thermo Fisher Scientific (1 mentions) Ique analyzer, by Intellicyt (1 mentions)

Close spelling variants of this product

Sulfo-NHS-LC-Biotin kit EZ-link biotinylation Kit (NHS-sulfo-LC-Biotin, Sulfo-NHS-LC-Biotinylation Kit EZ-Link Sulfo-NHS-LC-Biotinylation kit EZ Sulfo-NHS-LC-Biotinylation kit EZ-Link Sulfo-NHS-LC-Biotin kit EZ-Link Sulfo-NHS-LC-LC biotinylation kit EZ-Link Micro Sulfo-NHS-LC biotinylation kit EZ-Link™ Sulfo-NHS-LC-LC-Biotin kit Sulfo-NHS

8 protocols using nhs sulfo lc lc kit

1

Biotin Labeling of Antigens

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Antigens were biotinylated using the NHS-Sulfo-LC-LC kit following manufacturer’s instructions (ThermoFisher). Excess biotin was removed via size exclusion chromatography by using Zeba-Spin desalting columns (7 kDa cutoff, ThermoFisher).
Bartsch Y.C., Chen J.W., Kang J., Burns M.D., St Denis K.J., Sheehan M.L., Davis J.P., Edlow A.G., Balazs A.B., Yonker L.M, & Alter G. (2022). BNT162b2 induces robust cross-variant SARS-CoV-2 immunity in children. NPJ Vaccines, 7, 158.
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2

Quantifying Neutrophil Phagocytosis Capacity

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Phagocytosis score of primary human neutrophils was determined as described before.30 (link) Antigens were biotinylated with NHS-Sulfo-LC-LC kit according to the manufacturer’s instruction (Thermo Fisher). Excessive biotin was removed by size exclusion chromatography using Zeba-Spin desalting columns (7 kDa cutoff, Thermo Fisher). Biotinylated antigens were coupled to fluorescent neutravidin beads (Thermo Fisher) and incubated with 1:10 diluted plasma. Primary cells were derived from Ammonium-Chloride-Potassium (ACK) buffer lysed whole blood from healthy donors and incubated with immune complexes for one hour at 37°C. For the Fc-receptor blocking experiments, isolated neutrophils were pre-incubated with 5 μg/ml of FcγR2a (CD32A, clone IV.3, Bio X Cell Cat# BE0224, RRID:AB_2687707) and FcγR3 (CD16, clone: LNK16, Bio-Rad, RRID:AB_324304) five minutes prior to addition of neutrophils to the immune complexes. Neutrophils were stained for surface CD66b (BioLegend Cat# 305112, RRID:AB_2563294) expression, fixed with 4% para-formaldehyde, and analyzed an iQue analyzer (IntelliCyt) (Figure S8).
Bartsch Y.C., Cizmeci D., Kang J., Gao H., Shi W., Chandrashekar A., Collier A.R., Chen B., Barouch D.H, & Alter G. (2023). Selective SARS-CoV2 BA.2 escape of antibody Fc/Fc-receptor interactions. iScience, 26(5), 106582.
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3

Recombinant HIV Env Antigen Preparation

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Recombinant HIV Env gp120 (clade AE), gp120 (clade A) antigens were purchased from Immune Technology Corp. (NY, USA), tetanus toxoid from MassBiologics (MA, USA). Gp140 (clade AE) was obtained from Duke University DHVI Protein Production Facility. If required by the assay, antigens were biotinylated with N-hydroxysuccinimide (NHS)-Sulfo-LC-LC kit according to the manufacturer’s instruction (Thermo Fisher, MA, USA). Excessive biotin was removed by size exclusion chromatography using Zeba-Spin desalting columns (7-kDa cutoff; Thermo Fisher).
Bartsch Y.C., Loos C., Rossignol E., Fajnzylber J.M., Yuan D., Avihingsanon A., Ubolyam S., Jupimai T., Hirschel B., Ananworanich J., Lauffenburger D.A., Li J.Z., Alter G, & Julg B. (2021). Viral Rebound Kinetics Correlate with Distinct HIV Antibody Features. mBio, 12(2), e00170-21.
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4

SARS-CoV-2 Spike Protein Antigens

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SARS-CoV-2 D614G or variants of concern Spike and RBD proteins, in addition to NTD, S1, nucleocapsid, HCoV-HKU1 Spike (HKU1), and HCoV-OC43 Spike (OC43) antigens, were expressed in mammalian HEK293 cells and purchased from Sino Biological. Influenza haemagglutinin (HA) was obtained from Sino Biological. NHS-Sulfo-LC-LC kit was used for antigen biotinylating, according to the manufacturer’s instruction (Thermo Fisher Scientific).
Nziza N., Deng Y., Wood L., Dhanoa N., Dulit-Greenberg N., Chen T., Kane A., Swank Z., Davis J., Demokritou M., Chitnis A.P., Fasano A., Edlow A.G., Jain N., Horwitz B., McNamara R., Julg B., Shreffler W.G., Alter G, & Yonker L.M. (2023). Humoral profiles of toddlers and young children following SARS-CoV-2 mRNA vaccination. Research Square.
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5

SARS-CoV-2 Spike and Antigen Expression

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SARS-CoV-2 D614G or variants of concern Spike and RBD proteins, in addition to NTD, S1, nucleocapsid, HCoV-HKU1 Spike (HKU1), and HCoV-OC43 Spike (OC43) antigens, were expressed in mammalian HEK293 cells and purchased from Sino Biological. Influenza haemagglutinin (HA) was obtained from Sino Biological. NHS-Sulfo-LC-LC kit was used for antigen biotinylating, according to the manufacturer’s instruction (Thermo Fisher Scientific).
Nziza N., Deng Y., Wood L., Dhanoa N., Dulit-Greenberg N., Chen T., Kane A.S., Swank Z., Davis J.P., Demokritou M., Chitnis A.P., Fasano A., Edlow A.G., Jain N., Horwitz B.H., McNamara R.P., Walt D.R., Lauffenburger D.A., Julg B., Shreffler W.G., Alter G, & Yonker L.M. (2024). Humoral profiles of toddlers and young children following SARS-CoV-2 mRNA vaccination. Nature Communications, 15, 905.
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6

SARS-CoV-2 Spike Protein Production

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SARS-CoV-2 D614G or variants of concern Spike and RDB proteins of were expressed in mammalian HEK293 cells and obtained from SinoBiological. All antigens were biotinylated using an NHS-Sulfo-LC-LC kit according to the manufacturer’s instruction (Thermo Fisher Scientific). If required by the assay, excessive biotin was removed by size exclusion chromatography using Zeba-Spin desalting columns (7kDa cutoff, Thermo Fisher Scientific).
Bartsch Y.C., St Denis K.J., Kaplonek P., Kang J., Lam E.C., Burns M.D., Farkas E.J., Davis J.P., Boribong B.P., Edlow A.G., Fasano A., Shreffler W.G., Zavadska D., Johnson M., Goldblatt D., Balazs A.B., Yonker L.M, & Alter G. (2022). SARS-CoV-2 mRNA vaccination elicits robust antibody responses in children. Science Translational Medicine.
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7

Biotinylation and Purification Protocol

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All antigens were biotinylated using an NHS-Sulfo-LC-LC kit according to the manufacturer’s instruction (Thermo Fisher, MA, USA) if required by the assay and excessive biotin was removed by size exclusion chromatography using Zeba-Spin desalting columns (7kDa cutoff, Thermo Fisher).
Bartsch Y.C., St Denis K.J., Kaplonek P., Kang J., Lam E.C., Burns M.D., Farkas E.J., Davis J.P., Boribong B.P., Edlow A.G., Fasano A., Shreffler W., Zavadska D., Johnson M., Goldblatt D., Balazs A.B., Yonker L.M, & Alter G. (2022). Comprehensive antibody profiling of mRNA vaccination in children. bioRxiv.
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8

Preparation of SARS-CoV-2 Spike Variants

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Spike protein antigens for the D614G wildtype, alpha (B.1.1.7), beta (B.1.351), and delta (B.1.617.2) VOCs were obtained from Sino-Biologicals. Omicron (B.1.1.529) BA1 and BA2 Spikes were produced in house.39 (link) All antigens were produced in mammalian HEK293 cells. A strep-tag for purification was added to the C-terminus of the Omicron Spikes, whereas all other Spike variants had a His-tag at the C-terminus. All Spike antigens were expressed in the HexaPro (S-2P) form to stabilize the prefusion state of the protein. For functional assays all antigens were biotinylated using an NHS-Sulfo-LC-LC kit according to the manufacturer’s instruction (Thermo Fisher Scientific). Excessive biotin was removed by size exclusion chromatography using Zeba-Spin desalting columns (7kDa cutoff, Thermo Fisher Scientific).
Bartsch Y.C., Cizmeci D., Kang J., Gao H., Shi W., Chandrashekar A., Collier A.R., Chen B., Barouch D.H, & Alter G. (2023). Selective SARS-CoV2 BA.2 escape of antibody Fc/Fc-receptor interactions. iScience, 26(5), 106582.
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