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Streptavidin bb515

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Streptavidin-BB515 is a fluorescently-labeled streptavidin protein. Streptavidin has a high affinity for biotin, allowing it to be used as a detection reagent in various bioanalytical applications.

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Lab products found in correlation

Streptavidin pe, by BD (5 mentions) Biotin protein ligase bira kit, by Avidity (5 mentions) Streptavidin af647, by BioLegend (5 mentions) Ez link sulfo nhs lc biotinylation kit, by Thermo Fisher Scientific (5 mentions) Streptavidin bv711, by BD (5 mentions) Foxp3 transcription factor buffer set, by Thermo Fisher Scientific (2 mentions) Fc block 2.4g2, by BD (2 mentions) Alexa fluor 647 tagged ova, by Thermo Fisher Scientific (2 mentions) Np phycoerytherin pe, by Biosearch Technologies (2 mentions) Streptavidin bv421, by BioLegend (2 mentions) Cd326 magnetic beads, by Miltenyi Biotec (1 mentions)

Close spelling variants of this product

BD Horizon BB515 Streptavidin (2 citations)

8 protocols using streptavidin bb515

1

Multiparametric Flow Cytometry Analysis

Cited 1 time
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Spleens were forced through a 70-μm strainer into PBS supplemented with 5% fetal calfserum (FCS). Red blood cells(RBCs)werelysed withACKbuffer(Gilbco) followed by 10-min incubation with 1 μg/mL Fc Block (24G2; BD Biosciences), and then stained for 30 min at 4°C with the following antibodies acquired from Biolegend or BD Biosciences: B220 (RA3-6B2), CD138 (281-2), CD19 (ID3), CD38 (90), CD4 (RM4-5), CD86 (GL-1), CXCR5 (2G8), CXCR4 (2B11), Foxp3 (MF-23), GL-7 (GL7), ICOS (7E.17G9), IgD (11-26c.2a), IgG1 (A85-1), and PD-1 (J43). Antigen-specific GC responses were detected with Alexa Fluor 647-tagged OVA (Thermo Fisher), NP-phycoerytherin (PE) (Biosearch Technologies), or with FITC-tagged CRM (made in house) amplified by biotinylated α-FITC (Biolegend) and BB515 streptavidin (BD Biosciences). Phospho-S6 (pS6) staining was performed as described by Ersching et al. (2017) (link). For intracellular staining, samples were fixed and permeabilized prior to staining (Foxp3/transcription factor buffer set; eBioscience). Acquisition was conducted on a Cytek DxP11 FACSCalibur and analyzed on FlowJo X (Tree Star).
Silva M., Nguyen T.H., Philbrook P., Chu M., Sears O., Hatfield S., Abbott R.K., Kelsoe G, & Sitkovsky M.V. (2017). Targeted Elimination of Immunodominant B Cells Drives the Germinal Center Reaction toward Subdominant Epitopes. Cell reports, 21(13), 3672-3680.
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2

Multiparametric Flow Cytometry Analysis

Cited 1 time
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Spleens were forced through a 70-μm strainer into PBS supplemented with 5% fetal calfserum (FCS). Red blood cells(RBCs)werelysed withACKbuffer(Gilbco) followed by 10-min incubation with 1 μg/mL Fc Block (24G2; BD Biosciences), and then stained for 30 min at 4°C with the following antibodies acquired from Biolegend or BD Biosciences: B220 (RA3-6B2), CD138 (281-2), CD19 (ID3), CD38 (90), CD4 (RM4-5), CD86 (GL-1), CXCR5 (2G8), CXCR4 (2B11), Foxp3 (MF-23), GL-7 (GL7), ICOS (7E.17G9), IgD (11-26c.2a), IgG1 (A85-1), and PD-1 (J43). Antigen-specific GC responses were detected with Alexa Fluor 647-tagged OVA (Thermo Fisher), NP-phycoerytherin (PE) (Biosearch Technologies), or with FITC-tagged CRM (made in house) amplified by biotinylated α-FITC (Biolegend) and BB515 streptavidin (BD Biosciences). Phospho-S6 (pS6) staining was performed as described by Ersching et al. (2017) (link). For intracellular staining, samples were fixed and permeabilized prior to staining (Foxp3/transcription factor buffer set; eBioscience). Acquisition was conducted on a Cytek DxP11 FACSCalibur and analyzed on FlowJo X (Tree Star).
Silva M., Nguyen T.H., Philbrook P., Chu M., Sears O., Hatfield S., Abbott R.K., Kelsoe G, & Sitkovsky M.V. (2017). Targeted Elimination of Immunodominant B Cells Drives the Germinal Center Reaction toward Subdominant Epitopes. Cell reports, 21(13), 3672-3680.
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3

SARS-CoV-2 RBD and S Protein Biotinylation

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Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD or S were biotinylated using the Biotin-Protein Ligase-BIRA kit according to manufacturer’s instructions (Avidity) as described before5 (link). Ovalbumin (Sigma, A5503–1G) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BV711 for single-cell sorts (BD biosciences, 563262) or to streptavidin-BB515 for phenotyping panel (BD biosciences, 564453). RBD was conjugated to streptavidin-PE (BD Biosciences, 554061) and streptavidin-AF647 (Biolegend, 405237)5 (link),11 (link).
Schaefer-Babajew D., Wang Z., Muecksch F., Cho A., Raspe R., Johnson B., Canis M., DaSilva J., Ramos V., Turroja M., Millard K.G., Schmidt F., Dizon J., Shimelovich I., Yao K.H., Oliveira T.Y., Gazumyan A., Gaebler C., Bieniasz P.D., Hatziioannou T., Caskey M, & Nussenzweig M.C. (2022). Antibody feedback regulation of memory B cell development in SARS-CoV-2 mRNA vaccination. medRxiv.
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4

SARS-CoV-2 RBD and Ovalbumin Biotinylation

Cited 1 time
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Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD was biotinylated using the Biotin-Protein Ligase-BIRA kit according to manufacturer’s instructions (Avidity) as described before16 (link). Ovalbumin (Sigma, A5503-1G) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BV711 for single-cell sorts (BD biosciences, 563262) or to streptavidin-BB515 for phenotyping panel (BD biosciences, 564453). RBD was conjugated to streptavidin-PE (BD Biosciences, 554061) and streptavidin-AF647 (Biolegend, 405237)16 (link).
Muecksch F., Wang Z., Cho A., Gaebler C., Ben Tanfous T., DaSilva J., Bednarski E., Ramos V., Zong S., Johnson B., Raspe R., Schaefer-Babajew D., Shimeliovich I., Daga M., Yao K.H., Schmidt F., Millard K.G., Turroja M., Jankovic M., Oliveira T.Y., Gazumyan A., Caskey M., Hatziioannou T., Bieniasz P.D, & Nussenzweig M.C. (2022). Increased memory B cell potency and breadth after a SARS-CoV-2 mRNA boost. Nature, 607(7917), 128-134.
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5

Magnetic Bead-Based Enrichment of NCI-H2009 Cells

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NCI-H2009 cells were collected, washed with phosphate buffer saline (PBS), and
then incubated with or without 100 µl CD326 magnetic beads (Miltenyi Biotec
Inc., Bergisch Gladbach, Germany) at 4°C for 30 minutes. After washing twice,
the cells were incubated with anti-CD326 monoclonal antibodies CD326-FITC (3H)
(prepared in-house), CD326-AF488 (9C4), CD326-Biotin (9C4) (Biolegend, San
Diego, CA, USA), and CD326-Viobright 515 (Miltenyi Biotec Inc.),
streptavidin-BB515 (BD Biosciences, San Jose, CA, USA) and anti-mouse Dylight
488 polyclonal antibody (pAb) (Ab98757, Abcam, Cambridge, MA, USA) at 1 µg each
for 30 minutes at 4°C. Cells were washed twice, centrifuged at 300 × g for 5
minutes, and observed under a fluorescence microscope.
Wang H., Wu J., Zhang Q., Hao J., Wang Y., Li Z., Niu H., Zhang H, & Zhang S. (2021). A Modified Method to Isolate Circulating Tumor Cells and Identify by a Panel of Gene Mutations in Lung Cancer. Technology in Cancer Research & Treatment, 20, 1533033821995275.
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6

Biotinylation and Conjugation of SARS-CoV-2 Proteins

Cited 1 time
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Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD or S were biotinylated using the Biotin-Protein Ligase-BIRA kit according to manufacturer’s instructions (Avidity) as described previously5 (link). Ovalbumin (Sigma-Aldrich, A5503-1G) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BV711 for human single-cell sorts (BD Biosciences, 563262) or to streptavidin-BB515 for phenotyping (BD Biosciences, 564453). For all human and mouse experiments, RBD was conjugated to streptavidin-PE (BD Biosciences, 554061) and streptavidin-AF647 (BioLegend, 405237)5 (link),9 .
Schaefer-Babajew D., Wang Z., Muecksch F., Cho A., Loewe M., Cipolla M., Raspe R., Johnson B., Canis M., DaSilva J., Ramos V., Turroja M., Millard K.G., Schmidt F., Witte L., Dizon J., Shimeliovich I., Yao K.H., Oliveira T.Y., Gazumyan A., Gaebler C., Bieniasz P.D., Hatziioannou T., Caskey M, & Nussenzweig M.C. (2022). Antibody feedback regulates immune memory after SARS-CoV-2 mRNA vaccination. Nature, 613(7945), 735-742.
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7

Biotinylation and Conjugation of SARS-CoV-2 Proteins

Cited 1 time
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Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD and NTD were biotinylated using the Biotin-Protein Ligase-BIRA kit according to the manufacturer’s instructions (Avidity) as described before (Robbiani et al., 2020 (link)). Ovalbumin (A5503-1G; Sigma-Aldrich) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BB515 (564453; BD). RBD was conjugated to streptavidin-PE (554061; BD Biosciences) and streptavidin-AF647 (405237; BioLegend; Robbiani et al., 2020 (link)). NTD was conjugated to streptavidin-BV421 (405225; Biolegend) and streptavidin-BV711 (563262; BD Biosciences).
Cho A., Muecksch F., Wang Z., Ben Tanfous T., DaSilva J., Raspe R., Johnson B., Bednarski E., Ramos V., Schaefer-Babajew D., Shimeliovich I., Dizon J.P., Yao K.H., Schmidt F., Millard K.G., Turroja M., Jankovic M., Oliveira T.Y., Gazumyan A., Gaebler C., Caskey M., Hatziioannou T., Bieniasz P.D, & Nussenzweig M.C. (2022). Antibody evolution to SARS-CoV-2 after single-dose Ad26.COV2.S vaccine in humans. The Journal of Experimental Medicine, 219(8), e20220732.
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8

SARS-CoV-2 RBD and NTD Biotinylation

Cited 1 time
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Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD and NTD were biotinylated using the Biotin-Protein Ligase-BIRA kit according to the manufacturer’s instructions (Avidity) as described before (Robbiani et al., 2020 (link)). Ovalbumin (A5503-1G; Sigma-Aldrich) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BB515 (564453; BD). RBD was conjugated to streptavidin-PE (554061; BD Biosciences) and streptavidin-AF647 (405237; Biolegend; Robbiani et al., 2020 (link)). NTD was conjugated to streptavidin-BV421 (405225; Biolegend) and streptavidin-BV711 (563262; BD Biosciences).
Wang Z., Muecksch F., Muenn F., Cho A., Zong S., Raspe R., Ramos V., Johnson B., Ben Tanfous T., DaSilva J., Bednarski E., Guzman-Cardozo C., Turroja M., Millard K.G., Tober-Lau P., Hillus D., Yao K.H., Shimeliovich I., Dizon J., Kaczynska A., Jankovic M., Gazumyan A., Oliveira T.Y., Caskey M., Bieniasz P.D., Hatziioannou T., Kurth F., Sander L.E., Nussenzweig M.C, & Gaebler C. (2022). Humoral immunity to SARS-CoV-2 elicited by combination COVID-19 vaccination regimens. The Journal of Experimental Medicine, 219(10), e20220826.
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