Thrombin kit

Manufactured by Merck Group

The Thrombin kit is a laboratory product manufactured by Merck Group. It is a reagent used in the determination of thrombin activity. The kit provides the necessary components for the quantitative measurement of thrombin levels.

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Lab products found in correlation

Dylight 650, by Thermo Fisher Scientific (2 mentions) Anti his ab, by Thermo Fisher Scientific (2 mentions) Streptavidin pe sa pe, by Agilent Technologies (2 mentions) Sa pe, by Thermo Fisher Scientific (2 mentions)

2 protocols using thrombin kit

Recombinant Malaria Antigen Production and Characterization

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Recombinant His-tagged merozoite surface protein-1 (MSP-1₄₂) (amino acids 1398–1754; Accession P13828) and apical merozoite antigen-1 (AMA-1) (amino acids 26–478; Accession AAC47193) proteins from P. yoelii 17X were produced in E. coli as previously described.^{45 (link),46 (link)} After purification and refolding the MSP-1₄₂ and AMA-1 proteins, the His-tag was cleaved using a Thrombin kit (Millipore Sigma, St. Louis, MO). Confirmation of the His-tag removal was confirmed by W. blot using an anti-His Ab (ThermoFisher). MSP-1₄₂ and AMA-1 were biotinylated and tetramerized with streptavidin-PE (SA-PE; Agilent, Santa Clara, CA) as previously described.⁴⁷ A decoy reagent to gate out the non-MSP-1₄₂ or AMA-1–specific B cells was constructed by conjugating SA-PE to DyLight 650 (ThermoFisher) followed by washing and removal of any unbound DyLight 650 and incubating with an excess of biotin as described.⁴⁷

Brown S.L., Bauer J.J., Lee J., Ntirandekura E, & Stumhofer J.S. (2022). IgM+ and IgM− memory B cells represent heterogeneous populations capable of producing class-switched antibodies and germinal center B cells upon re-challenge with P. yoelii.. Journal of leukocyte biology, 112(5), 1115-1135.

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Recombinant protein production and biotinylation

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Recombinant His-tagged merozoite surface protein-1 (MSP-142) (amino acids 1398-1754; Accession P13828) and apical merozoite antigen-1 (AMA-1) (amino acids 26-478; Accession AAC47193) proteins from P. yoelii 17X were produced in E. coli as previously described. 44, 45 After purification and refolding the MSP-142 and AMA-1 proteins, the His-tag was cleaved using a Thrombin kit (Millipore Sigma, St. Louis, MO). Confirmation of the His-tag removal was confirmed by W. blot using an anti-His Ab (ThermoFisher). MSP-142 and AMA-1 were biotinylated and tetramerized with streptavidin-PE (SA-PE; Agilent, Santa Clara, CA) as previously described. 46 A decoy reagent to gate out the non-MSP-142 or AMA-1-specific B cells was constructed by conjugating SA-PE to DyLight 650 (ThermoFisher) followed by washing and removal of any unbound DyLight 650 and incubating with an excess of biotin as described. 46

Brown S.L., Bauer J.J., Lee J., Ntirandekura E., & Stumhofer J.S. (2021). IgM⁺and IgM^-memory B cells represent heterogeneous populations capable of producing class-switched antibodies and germinal center B cells upon re-challenge withP. yoelii.

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