Sa bv605

Manufactured by BioLegend

The SA-BV605 is a streptavidin conjugate that can be used to detect and quantify biotinylated molecules in various applications. It provides a strong and stable interaction with biotin, enabling sensitive detection.

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Lab products found in correlation

Cd45 30 f11, by Cytek Biosciences (2 mentions) Sa 711, by BioLegend (2 mentions) Facscanto 2, by BD (2 mentions) Facsaria fusion, by BD (2 mentions) Cd103 2e7, by Thermo Fisher Scientific (2 mentions) Ccr7 4b12, by BioLegend (2 mentions) Ab1225, by Merck Group (2 mentions) Percp cy5.5 anti ifn γ, by Thermo Fisher Scientific (2 mentions) Pacificblue anti cd20, by BioLegend (2 mentions) Biotin pd 1, by Thermo Fisher Scientific (2 mentions) Efluor660 anti il 21, by Thermo Fisher Scientific (2 mentions) Anti cd45ra fitc, by Thermo Fisher Scientific (2 mentions) Anti t bet, by Thermo Fisher Scientific (1 mentions) Alexa fluor 647 anti cxcr5, by BD (1 mentions) Anti t bet, by BD (1 mentions) Alexa fluor 488 anti il 10, by Thermo Fisher Scientific (1 mentions) Anti p stat4, by BD (1 mentions) Anti pstat5, by BD (1 mentions) Anti pstat3, by BD (1 mentions) Anti pstat6, by BD (1 mentions)

6 protocols using sa bv605

Splenic and Germinal Center B Cell Isolation

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Splenic B cells used for HTGTS-Rep-seq were purified from unimmunized 5 to 8-wk-old mice by MACS® Microbeads according to the manufacturer’s protocol. In brief, spleens were dissected out from unimmunized mice, prepared into single-cell suspensions, and stained with anti-B220 Microbeads for 20 min at 4 °C. The splenic B cells were collected using the LS column and MACS™ Separator. GC B cells used for Rep-SHM-seq were purified from 8 to 12-wk-old mice after eOD-GT8 60mer immunization. GC B cells were sorted for the phenotype B220⁺ (BV711: BioLegend 103255), CD95⁺ (PE-Cy7: eBioscience 557653) and GL7⁺ (PE: BioLegend 144607). CD4-binding site-specific GC B cells for single-cell RT-PCR were further selected for the phenotype eOD-GT8 Fc⁺ and ∆eOD-GT8 Fc⁻. The eOD-GT8 Fc was conjugated with Alexa Fluor 647 fluorescence (Thermo Fisher Scientific A30009). The ∆eOD-GT8 Fc was conjugated with Biotin (Thermo Fisher Scientific A30010) and then stained with SA-BV605 (BioLegend 405229).

Luo S., Jing C., Ye A.Y., Kratochvil S., Cottrell C.A., Koo J.H., Chapdelaine Williams A., Francisco L.V., Batra H., Lamperti E., Kalyuzhniy O., Zhang Y., Barbieri A., Manis J.P., Haynes B.F., Schief W.R., Batista F.D., Tian M, & Alt F.W. (2022). Humanized V(D)J-rearranging and TdT-expressing mouse vaccine models with physiological HIV-1 broadly neutralizing antibody precursors. Proceedings of the National Academy of Sciences of the United States of America, 120(1), e2217883120.

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Multicolor Flow Cytometry Panel

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Flow cytometry and cell sorting were completed on a FACS CantoII or FACS Aria Fusion instrument (BD) and analyzed using FlowJo analysis software (Tree Star). Staining was performed at 4ºC in the presence of Fc block (2.4G2) in magnetic-activated cell-sorting (MACS) buffer (PBS + .5% BSA + 2mM EDTA). The following antibodies were used; from BD Biosciences: CD117 (2B8), CD135 (A2F10.1), Ly6C (AL-21), MHCI (AF6–88.5), CD4 (RM4–5), CD8α (53–6.7), CD8β (53–5.8), CD11b (M1/70), B220 (RA3–6B2), CD64 (X54–5/7.1), CD19 (1D3), CD95 (Jo2), CD3 (145–2C11), CD45 (30-F11); from Tonbo Biosciences: MHCII (M5/114.15.2), CD44 (IM7), CD45.1 (A20), CD45.2 (104), CD11c (N418); from Biolegend: SA-BV605, SA-711, CCR7 (4B12), CD103 (2E7), XCR1 (ZET), CD115 (AFS98), Ter119 (Ter-119), Ly6G (1A8), TCRβ (H57–597), CD3 (145–2C11), CD8 (53–6.7), CD4 (RMA4–5), CD44 (IM7), CD40 (1C10), CD16/32 (93); from eBiosciences: TCRVα2 (B20.1), CD45.1 (A20), CD90.1 (HIS51), CD90.2 (53–2.1), F4/80 (BM8); from Invitrogen: CD172α (P84), CD45 (30F11); from Millipore/sigma: rabbit anti-ovalbumin (AB1225).

Ferris S.T., Durai V., Wu R., Theisen D.J., Ward J.P., Bern M.D., Davidson JT I.V., Bagadia P., Liu T., Briseño C.G., Li L., Gillanders W.E., Wu G.F., Yokoyama W.M., Murphy T.L., Schreiber R.D, & Murphy K.M. (2020). cDC1 prime and are licensed by CD4 T cells to induce anti-tumour immunity. Nature, 584(7822), 624-629.

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Multiparametric Profiling of T Cell Subsets

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eFluor660-anti–IL-21, Alexa Fluor 488–anti–IL-10, PerCP-Cy5.5-anti–IFN-γ, FITC-anti-CD45RA, PE-anti-ICOS, and anti-Tbet, and biotin–PD-1 were from eBioscience. Alexa Fluor 488–anti-GATA3, Alexa Fluor 647–anti-CXCR5, anti-pSTAT4, anti-pSTAT5, anti-pSTAT6, APC-anti-CD10, APC-Cy7-anti-CD4, BV605-anti-IgG, BV421-CD40L, BV711-anti–IL-2, PE-anti-pSTAT1, anti-RORγt, and Bcl-6, PE-Cy7-anti-CD25, and anti-CD27, PerCP-Cy5.5-anti-CD127, anti-pSTAT3, and anti-Tbet, SA-PerCpCy5.5, and IFN-γ were obtained from BD. Pacific Blue–anti-CD20 and SA-BV605 were purchased from BioLegend. Recombinant human IL-12 was purchased from R&D Systems. TGF-β, IL-1β, IL-6, IL-21, and IL-23 were obtained from PeproTech. PGE2 was purchased from Sigma-Aldrich. Human IL-4 and IL-10 were provided by R. de Waal Malefyt (DNAX Research Institute, Palo Alto, CA).

Ma C.S., Wong N., Rao G., Nguyen A., Avery D.T., Payne K., Torpy J., O’Young P., Deenick E., Bustamante J., Puel A., Okada S., Kobayashi M., Martinez-Barricarte R., Elliott M., Sebnem Kilic S., El Baghdadi J., Minegishi Y., Bousfiha A., Robertson N., Hambleton S., Arkwright P.D., French M., Blincoe A.K., Hsu P., Campbell D.E., Stormon M.O., Wong M., Adelstein S., Fulcher D.A., Cook M.C., Stepensky P., Boztug K., Beier R., Ikincioğullari A., Ziegler J.B., Gray P., Picard C., Boisson-Dupuis S., Phan T.G., Grimbacher B., Warnatz K., Holland S.M., Uzel G., Casanova J.L, & Tangye S.G. (2016). Unique and shared signaling pathways cooperate to regulate the differentiation of human CD4+ T cells into distinct effector subsets. The Journal of Experimental Medicine, 213(8), 1589-1608.

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Multi-Parameter Flow Cytometry Protocol

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eFluor660-anti-IL-21, PerCP-Cy5.5-anti-IFNγ, FITC-anti-CD45RA, biotin-PD-1 were from eBiosciences. Alexa647-anti-CXCR5 and anti-pSTAT1, APC-anti-CD10, APC-Cy7-anti-CD4, BV605-anti-IgG, PE-anti-pSTAT3 and anti-CCR6, Pe-Cy7-anti-CD25 and anti-CD27, PerCpCy5.5-anti-CD127, biotin-anti-IgA, SA-PerCpCy5.5, and recombinant IFNγ were from Becton Dickinson. BV421-anti-CXCR3, Pacific Blue-anti-CD20 and SA-BV605 were from Biolegend.

Ma C.S., Wong N., Rao G., Avery D.T., Torpy J., Hambridge T., Bustamante J., Okada S., Stoddard J.L., Deenick E.K., Pelham S.J., Payne K., Boisson-Dupuis S., Puel A., Kobayashi M., Arkwright P.D., Kilic S.S., Baghdadi J.E., Nonoyama S., Minegishi Y., Mahdaviani S.A., Mansouri D., Bousfiha A., Blincoe A.K., French M.A., Hsu P., Campbell D.E., Stormon M.O., Wong M., Adelstein S., Smart J.M., Fulcher D.A., Cook M.C., Phan T.G., Stepensky P., Boztug K., Kansu A., Ikincioğullari A., Baumann U., Beier R., Roscioli T., Ziegler J.B., Gray P., Picard C., Grimbacher B., Warnatz K., Holland S.M., Casanova J.L., Uzel G, & Tangye S.G. (2015). Monogenic mutations differentially impact the quantity and quality of T follicular helper cells in human primary immunodeficiencies. The Journal of allergy and clinical immunology, 136(4), 993-1006.e1.

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UV-Mediated Peptide Exchange for MHC Multimers

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Soluble biotinylated Class-I MHC monomers containing a UV-cleavable peptide in their binding groove were produced in-house according to published protocols⁷⁷ (link)
^,⁷⁸ (link) and stored at −80°C. The UV-dependent peptide exchange (25 μg/mL UV-monomer and 50 μg/mL peptide diluted in PBS) was performed at a wavelength of 366 nm for 1 h at 4°C. 24 h after UV-exchange, the following streptavidin-tagged fluorochromes were added at optimized ratios to the pMonomer solution: SA-PE (Invitrogen), SA-APC (Invitrogen), SA-BV421 (Biolegend), SA-BV605 (Biolegend), SA-PE-Cy7 (Biolegend), SA-PE-CF594 (BD Biosciences), SA-PE-Cy5 (BD Biosciences), SA-APC-R700 (BD Biosciences), SA-BB790-P (BD Biosciences; prototype kindly provided by Bob Balderas). To block unoccupied binding sites, D-biotin (20 μM; Avidity) was added 24 h after multimerization. Plates were stored in the dark at 4°C until use.

Meyer S., Blaas I., Bollineni R.C., Delic-Sarac M., Tran T.T., Knetter C., Dai K.Z., Madssen T.S., Vaage J.T., Gustavsen A., Yang W., Nissen-Meyer L.S., Douvlataniotis K., Laos M., Nielsen M.M., Thiede B., Søraas A., Lund-Johansen F., Rustad E.H, & Olweus J. (2023). Prevalent and immunodominant CD8 T cell epitopes are conserved in SARS-CoV-2 variants. Cell Reports, 42(1), 111995.

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Multicolor Flow Cytometry Panel

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