Strep tactin pe

Manufactured by IBA Lifesciences

Sourced in Germany

Strep-Tactin PE is a resin-based chromatography medium used for the purification of proteins with a Strep-tag. It binds to the Strep-tag with high affinity and specificity, allowing for the efficient capture and isolation of tagged proteins from complex mixtures.

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Lab products found in correlation

Streptactin apc, by IBA Lifesciences (2 mentions) Cytoflex flow cytometer, by Beckman Coulter (1 mentions) Calcium sensor dye efluor 514, by Thermo Fisher Scientific (1 mentions) Facs aria fusion cytometer, by BD (1 mentions) Cd14 percpcy5, by Thermo Fisher Scientific (1 mentions) Cd19 pe cy7, by BioLegend (1 mentions) Propidium iodide, by Thermo Fisher Scientific (1 mentions) Igm af488, by BioLegend (1 mentions) Cd8 percp cy5, by BioLegend (1 mentions) Anti human cd3 percp cy5, by BioLegend (1 mentions) Cd20 pe cy7, by BioLegend (1 mentions) Cd16 percp cy5, by BioLegend (1 mentions) Igd bv421, by BioLegend (1 mentions) Macs human b cell isolation kit, by Miltenyi Biotec (1 mentions) Cd27 bv510, by BD (1 mentions) Strep tactin magnetic beads, by IBA Lifesciences (1 mentions)

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Strep-Tactin (14 citations)

4 protocols using strep tactin pe

Expamers Binding and Activation Kinetics

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To measure Expamers cell-binding kinetics, freshly isolated T cells were resuspended in FACS buffer (PBS from Thermo Fischer Scientific with 0.5% BSA from Carl Roth) and measured continuously using a CytoFLEX Flow Cytometer (Beckman Coulter). During the measurement pre-assembled anti-CD3 and anti-CD28 Fab fragments with fluorescent Strep-Tactin PE (IBA Lifesciences) were added to T cells. Increase in signal intensity was recorded.
Changes in intracellular Ca²⁺ were monitored using a flow cytometer LSRI analyzer (BD Biosciences). Cells were stained with Calcium Sensor Dye eFluor514 (eBiosciences) according to manufacturer’s instructions and illuminated with 325 nm laser line of a helium-cadmium laser. Fluorescence emissions from 390 to 420 nm and from 500 to 520 nm were detected simultaneously, and changes in the ratio of the two emission intensities were analyzed.
Residual content of individual Expamers components on T cell surfaces was measured using flow cytometric detection after 8 days of culture. For Fab fragment detection, T cells were stained with fluorescent Strep-Tactin PE (IBA Lifesciences) that should be able to bind to the Strep-tag of remaining Fabs. For detection of Strep-Tactin multimer backbone DyLight488 anti-streptavidin antibody (Vector Laboratories) was used.

Poltorak M.P., Graef P., Tschulik C., Wagner M., Cletiu V., Dreher S., Borjan B., Fraessle S.P., Effenberger M., Turk M., Busch D.H., Plitzko J., Kugler D.G., Ragan S., Schmidt T., Stemberger C, & Germeroth L. (2020). Expamers: a new technology to control T cell activation. Scientific Reports, 10, 17832.

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HBV-specific CD8+ T cell detection

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HBV-specific CD8⁺ T cells were detected through staining with MHC class I multimers conjugated with the H-2K^b-restricted HBV-derived peptides C_93–100 (C₉₃, MGLKFRQL) and S_190-197 (S₁₉₀, VWLSAIWM), as described previously.¹⁰ ^,²¹ (link) As a control, staining with multimer conjugated with the ovalbumin-derived peptide S8L₂₅₇ (OVA_S8L, SIINFEKL) was performed. Prior to use, C₉₃ and OVA_S8L multimers (kindly provided by Dirk Busch, Technical University of Munich, Germany) were labelled with Streptactin-PE (IBA Lifesciences, Göttingen, Germany), as previously described.¹⁰ ^,²¹ (link)

Sacherl J., Kosinska A.D., Kemter K., Kächele M., Laumen S.C., Kerth H.A., Öz E.A., Wolff L.S., Su J., Essbauer S., Sutter G., Scholz M., Singethan K., Altrichter J, & Protzer U. (2022). Efficient stabilization of therapeutic hepatitis B vaccine components by amino-acid formulation maintains its potential to break immune tolerance. JHEP Reports, 5(2), 100603.

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Tetramer-based Identification and Sorting of B Cells

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B cells were eluted from PBMCs using a MACS Human B Cell isolation kit (Miltenyi Biotec). B cells were stained with rGP38 (IbAr10200) that had been tetramerized at 25 nM using Streptactin-PE (IBA Lifesciences) and Streptactin-APC (IBA Lifesciences). B cells were simultaneously stained with rGP38-Streptactin-PE and rGP38-Streptactin-APC tetramers for 1 hour on ice. Cells were washed twice in buffer (PBS, FBS, EDTA). Next, B cells were stained with a panel of antibodies. Donor 1 PBMCs were stained with a cocktail of anti-human CD3 PerCP-Cy5.5 (Biolegend), CD8 PerCP-Cy5.5 (Biolegend), CD14 PerCP-Cy5.5 (Invitrogen), CD16 PerCP-Cy5.5 (Biolegend), propidium iodide (PI) (Invitrogen), CD19 PE-Cy7 (Biolegend), CD27 BV510 (BD Biosciences), IgM BV711 (BD Biosciences), IgD BV421 (Biolegend), IgG BV605 (BD Biosciences), and IgA AF488 (Abcam). Donor 5 and 6 PBMCs were stained with a cocktail of anti-human CD3 PerCP-Cy5.5 (Biolegend), CD8 PerCP-Cy5.5 (Biolegend), CD14 PerCP-Cy5.5 (Invitrogen), CD16 PerCP-Cy5.5 (Biolegend), PI (Invitrogen), CD19 PE-Cy7 (Biolegend), CD20 PE-Cy7 (Biolegend), CD27 BV510 (BD Biosciences), IgM AF488 (Biolegend), and IgD BV421 (Biolegend). B cells were washed twice in buffer and run on a FACS Aria Fusion Cytometer (BD Biosciences). B cells were sorted into Super Script III reaction buffer (ThermoFisher Scientific) in 96-well Costar plates and frozen at −80 °C.

Shin O.S., Monticelli S.R., Hjorth C.K., Hornet V., Doyle M., Abelson D., Kuehne A.I., Wang A., Bakken R.R., Mishra A., Middlecamp M., Champney E., Stuart L., Maurer D.P., Li J., Berrigan J., Barajas J., Balinandi S., Lutwama J.J., Lobel L., Zeitlin L., Walker L.M., Dye J.M., Chandran K., Herbert A.S., Pauli N.T, & McLellan J.S. (2024). Crimean-Congo Hemorrhagic Fever Survivors Elicit Protective Non-Neutralizing Antibodies that Target 11 Overlapping Regions on Viral Glycoprotein GP38. bioRxiv.

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Generation of MHC Streptamers for Cell Staining

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Generation of MHC Streptamers was performed as previously described [19 (link), 26 (link)]. In brief, inclusion body-derived recombinant MHC heavy chain was urea-denatured and refolded in the presence of peptide and β2 microglobulin. Complexes were purified by fast protein liquid chromatography (FPLC). Multimerization was performed with Strep-Tactin^® PE (cell staining; IBA Lifesciences, Goettingen, Germany), Strep-Tactin^® APC (cell staining; IBA Lifesciences) or Strep-Tactin^® magnetic beads (cell isolation; IBA Lifesciences). The following MHC Streptamers were used: HLA-B*07:02/pp65_417-427 (TPRVTGGGAM), HLA-C*07:02/IE-1_309−317 (CRVLCCYVL) and HLA-C*07:02/MAGE-A12_170-178 (VRIGHLYIL).

Schlott F., Steubl D., Ameres S., Moosmann A., Dreher S., Heemann U., Hösel V., Busch D.H, & Neuenhahn M. (2018). Characterization and clinical enrichment of HLA-C*07:02-restricted Cytomegalovirus-specific CD8+ T cells. PLoS ONE, 13(2), e0193554.

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