Streptactin apc

Manufactured by IBA Lifesciences

Sourced in Germany

Streptactin-APC is a lab equipment product used for the purification and detection of recombinant proteins. It functions as an affinity-based ligand for the purification of proteins tagged with Strep-tag II, a small peptide affinity tag. Streptactin-APC provides a reversible binding and elution system for efficient protein recovery.

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Lab products found in correlation

Strep tactin pe, by IBA Lifesciences (2 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) Sars cov 2 rbd mfc, by Sino Biological (1 mentions) Strep tactin magnetic beads, by IBA Lifesciences (1 mentions) Facscanto 2, by BD (1 mentions) Goat f ab 2 anti human kappa fitc, by Southern Biotech (1 mentions) Goat f ab 2 anti human lambda fitc, by Southern Biotech (1 mentions)

4 protocols using streptactin apc

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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Isolation of SARS-CoV-2 Reactive Cells

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Transfected cells were sorted for HDR + by staining for StrepTactin-APC 1:100 (IBA lifesciences, Cat: 6-5010-001) and anti-hIgG : AF488 1:100 (Jackson ImmunoResearch, Cat: 109-545-003). Enriched cells were then selected for antigen binding by incubating with SARS-CoV-2 S1-mFc 1:75 (Sinobiological, Cat: 40591-V05H1), S2-mFc 1:75 (Sinobiological, Cat: 40,590-V05B) on ice for 20 min, followed by a single wash and a secondary staining with anti-mFc IgG1: PE and/or APC [clone RMG1-1] at 1:100 (Biolegend, Cat: 406,608/406,610). After enrichment for S1, cells were also tested for binding to SARS-CoV-2 RBD-mFc 1:375 (Sinobiological, Cat: 40,592-V05H), and the same secondary staining. FACS was analyzed using FlowJo X software. At least 20′000 live events were acquired per sample to ensure sufficient statistical sampling.

Ehling R.A., Weber C.R., Mason D.M., Friedensohn S., Wagner B., Bieberich F., Kapetanovic E., Vazquez-Lombardi R., Di Roberto R.B., Hong K.L., Wagner C., Pataia M., Overath M.D., Sheward D.J., Murrell B., Yermanos A., Cuny A.P., Savic M., Rudolf F, & Reddy S.T. (2021). SARS-CoV-2 reactive and neutralizing antibodies discovered by single-cell sequencing of plasma cells and mammalian display. Cell Reports, 38(3), 110242.

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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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Competitive Binding Analysis of hMPV Antibodies

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Competitive binning experiments were performed by evaluating IgG binding to hMPV F in the presence and absence of pre-complexed competitor Fab, as previously described (Fels et al., 2021) (link). Briefly, hMPV preF A1 (25 nM) was complexed with competitor Fab (0.5 mM) for 30 min on ice. The antigen-Fab complex was then incubated for 5 min with yeast expressing anti-hMPV IgG. After washing with PBSF to remove unbound antigen, the bound antigen was detected using Strep-Tactin APC (IBA Lifesciences, catalog no. 6-5010-001) and antibody light chain was detected using Goat F(ab')2 anti-human kappa FITC (SouthernBiotech, catalog no. 2062-02) and Goat F(ab')2 anti-human lambda FITC (SouthernBiotech, catalog no. 2072-02) . The samples were analyzed by flow cytometry using a FACSCanto II (BD Biosciences). For mAbs that displayed detectable binding to hMPV preF A1 in the absence of a competitor Fab, competitive binding was evaluated based on the fold reduction in antigen binding observed in the presence of the competitor Fabs. Fold-reductions greater than 4-fold were classified as competitive binding.

Rappazzo C.G., Hsieh C.L., Rush S.A., Esterman E.S., Delgado T., Geoghegan J.C., Wec A.Z., Sakharkar M., Más V., McLellan J.S, & Walker L.M. (2022). Potently neutralizing and protective anti-human metapneumovirus antibodies target diverse sites on the fusion glycoprotein. Immunity, 55(9).

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