Streptavidin pe sa pe

Manufactured by Agilent Technologies

Streptavidin-PE (SA-PE) is a conjugate of streptavidin and the fluorescent dye phycoerythrin (PE). Streptavidin is a tetrameric protein that binds to biotin with high affinity. The PE dye provides a strong fluorescent signal for detection and analysis purposes.

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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) Sa pe, by Thermo Fisher Scientific (2 mentions) Thrombin kit, by Merck Group (2 mentions)

Spelling variants of this product

PE-Streptavidin (14 citations) SA-PE (4 citations)

2 protocols using streptavidin pe sa pe

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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