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Sulfo nhs lc biotinylation kit

Manufactured by Thermo Fisher Scientific
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Sourced in Canada

The Sulfo-NHS-LC-Biotinylation Kit is a laboratory product that facilitates the biotinylation of proteins. It contains the reagent Sulfo-NHS-LC-Biotin, which is used to covalently attach a biotin label to primary amines on proteins. This allows for the detection and analysis of proteins through biotin-avidin interactions.

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Lab products found in correlation

Nanp 9 peptide, by Biomatik (2 mentions) Premium grade sa pe, by Thermo Fisher Scientific (2 mentions) Sa apc, by Thermo Fisher Scientific (2 mentions) Fortessa flow cytometer, by BD (2 mentions) Decorin, by Merck Group (1 mentions) Series s sensor chip cm5, by GE Healthcare (1 mentions) Neutravidin protein, by Thermo Fisher Scientific (1 mentions) Biacore t200 system, by GE Healthcare (1 mentions)

Close spelling variants of this product

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

5 protocols using sulfo nhs lc biotinylation kit

1

Multicolor Flow Cytometry for Malaria Antigen Detection

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PfPfMSP1, PfPfAMA1 and TT were biotinylated with the Sulfo-NHS-LC-Biotinylation Kit (ThermoFisher) at a ratio of 1:1 according to the manufacturer’s instructions. For detection of PfCSP-specific cells biotinylated (NANP)9 peptide was sourced from Biomatik (Ontario, Canada). Biotinylated Ags were incubated with premium-grade SA-PE and SA-APC (Molecular Probes) at a molar ratio of 4:1, added four times with 15 min incubation at room temperature.
PBMCs, processed and cryopreserved at 5 × 106 [31 (link)], were thawed and washed in PBS with 2% heat inactivated FBS. Approximately 2.5 × 106 cells were incubated with 1 μL of both SA-PE and SA-APC tetramers for 20 min at room temperature. Cells were further incubated for an additional 30 min with human B-cell surface molecules using the antibodies listed in Supporting Information Table 2. One million events were acquired on a BD Fortessa flow cytometer (BD Biosciences, San Jose, CA) and the data analysed using FlowJo software (Tree Star, Inc, Ashland, OR).
Aye R., Sutton H.J., Nduati E.W., Kai O., Mwacharo J., Musyoki J., Otieno E., Wambua J., Bejon P., Cockburn I.A, & Ndungu F.M. (2020). Malaria exposure drives both cognate and bystander human B cells to adopt an atypical phenotype. European journal of immunology, 50(8), 1187-1194.
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2

Quantifying VAR2CSA-CSPG Binding Affinity

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The binding affinity of the VAR2CSA recombinant proteins to CSPG was measured using a Biacore T200 system (GE Healthcare). Briefly, decorin (Sigma) was biotinylated non-specifically at lysine residues in the protein core using Sulfo-NHS-LC-Biotinylation Kit (Thermo). NeutrAvidin protein (Thermo Scientific) was covalently immobilized on a Series S sensor chip CM5 (GE Healthcare) using N-hydroxysuccinimide (NHS) amine coupling chemistry, in flow cell 4 (Fc-4), while a biotin-labeled Pfs25 recombinant was similarly immobilized in the reference Fc-3. Protein binding affinity to the immobilized CSPG was obtained by subtracting the response on reference Fc-3 from that on Fc-4, after injection of a serial dilution of the full-length VAR2CSA recombinants. The affinity analysis was performed in Biacore T200 evaluation software (version 3.2) and the steady-state model was used to calculate the dissociation constant KD.
Renn J.P., Doritchamou J.Y., Tentokam B.C., Morrison R.D., Cowles M.V., Burkhardt M., Ma R., Mahamar A., Attaher O., Diarra B.S., Traore M., Dicko A., Tolia N.H., Fried M, & Duffy P.E. (2021). Allelic variants of full-length VAR2CSA, the placental malaria vaccine candidate, differ in antigenicity and receptor binding affinity. Communications Biology, 4, 1309.
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3

Induction and Detection of Malaria Immunity

Cited 1 time
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For induction of GC reactions, each mouse was intravenously injected with 200 μl of the SRBC suspension containing 2 × 108 SRBCs, and spleens were analyzed at day 10. Alternatively, Plasmodium berghei parasites engineered to express Plasmodium falciparum CSP (PfCSP) in place of the endogenous P. berghei CSP molecule (Pb-PfSPZ) (51 (link)) were used. Parasites were maintained by serial passage through Anopheles stephensi mosquitoes. Mice were immunized intravenously with 5 × 104 irradiated (15 kRad) Pb-PfSPZ dissected by hand from the salivary glands of A. stephensi mosquitoes as described previously.
For the detection of PfCSP-specific cells, a nine-times repeat of asparagine-alanine-asparagine-proline (NANP)9 peptide was sourced from Biomatik (Ontario, Canada) and was biotinylated with the Sulfo-NHS-LC Biotinylation Kit (Thermo Fisher Scientific) at a ratio of 1:1 according to the manufacturer’s instructions. Biotinylated antigens were incubated with premium-grade streptavidin-phycoerythrin and streptavidin-allophycocyanine (Molecular Probes) at a molar ratio of 4:1 and added four times with 15-min incubation at room temperature.
A multimorphic mutation in IRF4 causes human autosomal dominant combined immunodeficiency. (2023). Science immunology, 8(79), eade7953.
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4

Multicolor Flow Cytometry for Malaria Antigen Detection

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PfPfMSP1, PfPfAMA1 and TT were biotinylated with the Sulfo-NHS-LC-Biotinylation Kit (ThermoFisher) at a ratio of 1:1 according to the manufacturer’s instructions. For detection of PfCSP-specific cells biotinylated (NANP)9 peptide was sourced from Biomatik (Ontario, Canada). Biotinylated Ags were incubated with premium-grade SA-PE and SA-APC (Molecular Probes) at a molar ratio of 4:1, added four times with 15 min incubation at room temperature.
PBMCs, processed and cryopreserved at 5 × 106 [31 (link)], were thawed and washed in PBS with 2% heat inactivated FBS. Approximately 2.5 × 106 cells were incubated with 1 μL of both SA-PE and SA-APC tetramers for 20 min at room temperature. Cells were further incubated for an additional 30 min with human B-cell surface molecules using the antibodies listed in Supporting Information Table 2. One million events were acquired on a BD Fortessa flow cytometer (BD Biosciences, San Jose, CA) and the data analysed using FlowJo software (Tree Star, Inc, Ashland, OR).
Aye R., Sutton H.J., Nduati E.W., Kai O., Mwacharo J., Musyoki J., Otieno E., Wambua J., Bejon P., Cockburn I.A, & Ndungu F.M. (2020). Malaria exposure drives both cognate and bystander human B cells to adopt an atypical phenotype. European journal of immunology, 50(8), 1187-1194.
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5

Biotinylation and Fluorescent Labeling

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Pf MSP1, AMA1 and TT were biotinylated with the Sulfo-NHS-LC-Biotinylation Kit (ThermoFisher) at a ratio of 1:1 according to the manufacturer’s instructions, biotinylated (NANP)9 repeat region of Pf CSP was sourced from Biomatik (Ontario, Canada). Biotinylated antigens were incubated with premium-grade SA-PE and SA-APC (Molecular Probes) or SA-BV421 and SA-BB660 (Biolegend and BD Horizon) at a molar ratio of 4:1, added four times with 15 min incubation at room temperature.
Sutton H.J., Aye R., Idris A.H., Vistein R., Nduati E., Kai O., Mwacharo J., Li X., Gao X., Andrews T.D., Koutsakos M., Nguyen T.H., Nekrasov M., Milburn P., Eltahla A., Berry A.A., KC N., Chakravarty S., Sim B.K., Wheatley A.K., Kent S.J., Hoffman S.L., Lyke K.E., Bejon P., Luciani F., Kedzierska K., Seder R.A., Ndungu F.M, & Cockburn I.A. (2021). Atypical B cells are part of an alternative lineage of B cells that participates in responses to vaccination and infection in humans. Cell Reports, 34(6), 108684.
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