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Apc conjugated streptavidin

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APC-conjugated streptavidin is a fluorescent-labeled protein that binds strongly and specifically to biotin. It is commonly used in flow cytometry, immunoassays, and other applications requiring the detection or separation of biotinylated molecules.

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Lab products found in correlation

Lsrfortessa, by BD (2 mentions) Facscalibur, by BD (1 mentions) Sy3200, by Sony (1 mentions) Cytofix cytoperm solution, by BD (1 mentions) Pe conjugated anti hcd25 clone bc96, by BioLegend (1 mentions) Apc conjugatedanti hcd25 clone bc96, by BioLegend (1 mentions) Arthrobacter ureafaciens, by Merck Group (1 mentions) Facscanto 2, by BD (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions) Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions) Percp cy5, by Thermo Fisher Scientific (1 mentions) Percp cy5, by BD (1 mentions) Ammonium chloride, by Merck Group (1 mentions) Ctla 4, by BioLegend (1 mentions) Siglec f, by BioLegend (1 mentions) Fcεri, by BioLegend (1 mentions) Cd278, by Thermo Fisher Scientific (1 mentions) Cd11b, by BioLegend (1 mentions) F4 80, by BioLegend (1 mentions)

Spelling variants of this product

Streptavidin-APC (121 citations) APC-Cy7-conjugated streptavidin (7 citations) Streptavidin conjugated PE or APC (2 citations) APC-eFluor 780-conjugated streptavidin, (2 citations) Allophycocyanin (APC)–conjugated streptavidin (2 citations)

22 protocols using apc conjugated streptavidin

1

Flow Cytometry Analyses and Sorting

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Flow cytometric analyses were performed on a FACSCalibur or BD LSRFortessa (BD). Sorting was conducted on a BD Aria (user-operated) or on a Sony Sy3200 through the Siteman Cancer Center Flow Cytometry Core Facility. FITC-conjugated anti-CD45R/B220 (clone RA3-6B2), phycoerythrin (PE)-conjugated anti-CD43 (clone S7), FITC-conjugated anti-CD43 (clone S7), PE-Cy7–conjugated anti-CD45/B220 (clone RA3-6B2), allophycocyanin (APC)-conjugated anti-IgM (clone II/41), PE-conjugated anti-CD40 (clone 1C10), and biotin-conjugated anti-CD40 (clone 3/23) were purchased from BD. APC-conjugated streptavidin was purchased from eBioscience. PE-conjugated anti-hCD25 (clone BC96) and APC-conjugated anti-hCD25 (clone BC96) were purchased from BioLegend. APC-conjugated anti-mCherry (clone 16D7) was purchased from Life Technologies. Staining for intracellular proteins was performed using Cytofix/Cytoperm solution (BD).
Bednarski J.J., Pandey R., Schulte E., White L.S., Chen B.R., Sandoval G.J., Kohyama M., Haldar M., Nickless A., Trott A., Cheng G., Murphy K.M., Bassing C.H., Payton J.E, & Sleckman B.P. (2016). RAG-mediated DNA double-strand breaks activate a cell type–specific checkpoint to inhibit pre–B cell receptor signals. The Journal of Experimental Medicine, 213(2), 209-223.
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2

Sialic Acid and EGFR Targeting Agents

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The α2,3-linked sialyl residues on cells were stained with 5 μg/mL biotinylated Maackia Amurensis Lectin II (MAL II, Vector laboratories) and detected with APC-conjugated Streptavidin (eBioscience). Neuraminidase Clostridium Perfingens (Sigma Aldrich), Vibrio Cholerae (Merck) Arthrobacter ureafaciens (Sigma Aldrich), and Salmonella Typhimurium (SIALST) were used at 0.1 U/mL to hydrolyze terminal sialic acid. Surface EGFR expression was determined with an anti-EGFR antibody (clone AY13, Biolegend). In the CD47 binding competition assay CD47-PE (CC2C6, Biolegend) was used to detect displacement of the SIRPα fusion protein. Detection of Siglec ligands on tumor cells was performed using 10 μg/mL recombinant human Siglec-5, -7, and -9 Fc chimera protein (Bio-Techne). IgA3.0 antibodies, hereafter called IgA, against EGFR (Cetuximab), EpCAM (HeING), and HER2 (Trastuzumab) were produced and purified in house as described in Stip et al. (2023, in press). CD47-blocking, SIRPα fusion protein was produced and purified in house as described in Chernyavska et al. (2022) [28 (link)]. IgG2 FcKO Siglec-9 blocking antibody (mAbA) was produced and purified in house [23 (link)].
Chan C., Lustig M., Jansen J.H., Garcia Villagrasa L., Raymakers L., Daamen L.A., Valerius T., van Tetering G, & Leusen J.H. (2023). Sialic Acids on Tumor Cells Modulate IgA Therapy by Neutrophils via Inhibitory Receptors Siglec-7 and Siglec-9. Cancers, 15(13), 3405.
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3

Comprehensive Mouse Spleen and Bone Marrow Immunophenotyping

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Briefly, mouse spleens and BM were collected and dissociated through a 40μm strainer to produce a single cell suspension. Red blood cells were lysed using 0.86% ammonium chloride (Sigma). Cells were counted, washed in flow cytometry buffer (1% BSA (Sigma), 2mM EDTA (Invitrogen) in PBS. FcR were blocked with 2.4G2 (BioXCell) and Rat IgG (Invitrogen) while staining with McAb at 40 for 30 minutes. Intracellular staining was carried out using the eBioscience kit. Samples were acquired using a FACSCanto II or LSRFortessa flow cytometer (BD Biosciences) and analyzed with FlowJo software (Tree Star, Inc.). The following mAb against mouse antigens were used as FITC, PE, PerCP-Cy5.5, PE-Cy7, allophycocyanin (APC), APC-ef780, Pacific blue, AF450, AF700, PE Texas Red, or biotin conjugates: FcεRI, AA4.1, CD23, CD43, IgM, FcγRII/III, ckit, Sca-1, CD34, CD138, CD45R (B220; RA3-6B2), CD278 (ICOS; C398.4A), IgD (11–26) (eBioscience), CD4 (RM4–5), CD8a (53–6.7), CD95 (Fas; Jo2), CXCR5 (2G8) (BD Biosciences), CD3 (17A2), CD19 (6D5), Foxp3, CTLA-4, Siglec F, CD11b, F4/80, TCRβ, Gr-1, FcεRI, and CD279 (PD-1; J43) (Biolegend). Biotinylated antibodies were detected using PerCP-Cy5.5– (BD Biosciences) or APC-conjugated streptavidin (eBioscience). FITC or biotin-conjugated PNA was obtained from Vector Laboratories. Plots shown are on a Logicle scale.
Zaretsky A.G., Konradt C., Dépis F., Wing J.B., Goenka R., Atria D.G., Silver J.S., Cho S., Wolf A.I., Quinn W.J., Engiles J.B., Brown D.C., Beiting D., Erikson J., Allman D., Cancro M.P., Sakaguchi S., Lu L., Benoist C.O, & Hunter C.A. (2017). T Regulatory Cells Support Plasma Cell Populations in the Bone Marrow. Cell reports, 18(8), 1906-1916.
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4

Antigen Uptake and Presentation Assay

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In vitro assay was performed with immature BMDCs at day 6 treated with EαGFP peptide (50 µg/mL) for 4 h at 37 °C with 5% of CO2. BMDCs were then collected, washed twice with 2% FBS, 2 µM EDTA PBS (MACS buffer) and stained with biotin-conjugated anti-mouse Eα 52–68 peptide (eBioscience, Cat#13-5741-81), for 30 min at 4 °C. Cells were washed twice with 2% FBS, 2 µM EDTA PBS (MACS buffer) and stained with APC-conjugated streptavidin (eBioscience, Cat#17-4317-82) for 30 min at 4 °C. Antigen uptake and presentation were assessed by flow cytometry.
For in vivo assay, WT and apoE KO mice were injected i.p. with EαGFP peptide (20 µg/mL). After 24 h, peritoneal lavage was collected, washed in 2% FBS, 2 µM EDTA PBS (MACS buffer) and stained for flow cytometry analysis with eFluor660-CD11c (eBioscience, Cat#500114) and eFluor 450- MHCII (eBioscience, Cat#48-5321-82).
Bonacina F., Coe D., Wang G., Longhi M.P., Baragetti A., Moregola A., Garlaschelli K., Uboldi P., Pellegatta F., Grigore L., Da Dalt L., Annoni A., Gregori S., Xiao Q., Caruso D., Mitro N., Catapano A.L., Marelli-Berg F.M, & Norata G.D. (2018). Myeloid apolipoprotein E controls dendritic cell antigen presentation and T cell activation. Nature Communications, 9, 3083.
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5

Detecting LPS Binding on Monocytes

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Monocytes were stimulated with biotinylated LPS (10 μg ml−1, Invivogen) for the indicated times. Membrane-bound LPS was detected by incubation with AlexaFluor 450-conjugated streptavidin (1:200, eBioscience). After staining, cells were rinsed extensively with PBS to remove excess streptavidin, and then fixed and permeabilized with BD Cytofix/Cytoperm buffer. Intracellular LPS was stained using APC-conjugated streptavidin (1:200, eBioscience) and data were acquired on a LSR II flow cytometer (BD Biosciences) for analysis using FlowJo software (TreeStar).
Viganò E., Diamond C.E., Spreafico R., Balachander A., Sobota R.M, & Mortellaro A. (2015). Human caspase-4 and caspase-5 regulate the one-step non-canonical inflammasome activation in monocytes. Nature Communications, 6, 8761.
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6

Multicolor Flow Cytometry Analysis

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Cells were phenotypically analyzed using a multicolor flow cytometer Navios (Beckman Coulter, Mijdrecht, Netherlands). The following conjugated mAb were used: CD25/Pe-Cy7 (M-A251), HLA-DR/FITC (L243) (both from BD Bioscience); TIGIT/PE (MBSA43, eBioscience, Vienna, Austria), CD3/ECD (UCHT1), CD4/PE-Cy5.5 (1388.2), CD8/APC-AF700 (B9.11) (all from Beckman Coulter), TNFR2/APC (#22235; R&D), and Fixable Viability Dye eFluor780 (eBioscience). To detect the expression of mTNFα, cells were first stained with biotin-labelled Infliximab followed with APC-conjugated streptavidin (eBioscience). For intracellular staining, EZH2/PE (11/EZH2, BD Bioscience), FOXP3/eFluor 450 (PCH101), and Helios/AlexFluor 647 (22F6) (both from eBioscience) were used after fix-perm-treatment of cells, according to the manufacturer’s instructions. Isotype matched control antibodies were used to define marker settings. Data were analyzed using the software Kaluza (Beckman Coulter).
Urbano P.C., Koenen H.J., Joosten I, & He X. (2018). An Autocrine TNFα–Tumor Necrosis Factor Receptor 2 Loop Promotes Epigenetic Effects Inducing Human Treg Stability In Vitro. Frontiers in Immunology, 9, 573.
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7

Quantifying IL-2-Fc Binding on Immune Cells

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Red blood cell-depleted splenocytes from a FoxP3-DTR/GFP transgenic mouse were incubated with Fc-Block (anti-mouse CD16/CD32, BD), followed by washing in flow cytometry buffer and staining with fluorophore-conjugated anti-CD3e, anti-CD8a, anti-CD4, anti-CD44 and anti-NK1.1 antibodies. Stained cells were washed, seeded in a 96-well plate (1 × 106 cells per well) and re-suspended in serially diluted IL-2WTFc-biotin. After incubation for 30 min on ice, cells were stained with APC-conjugated streptavidin (eBioscience). The levels of IL-2WTFc bound to the surface of MP CD8 (CD3+ CD8+ CD44high), NK cells (CD3− NK1.1+) and Tregs (CD3+ CD4+ FoxP3+) were determined by flow cytometry.
Vazquez-Lombardi R., Loetsch C., Zinkl D., Jackson J., Schofield P., Deenick E.K., King C., Phan T.G., Webster K.E., Sprent J, & Christ D. (2017). Potent antitumour activity of interleukin-2-Fc fusion proteins requires Fc-mediated depletion of regulatory T-cells. Nature Communications, 8, 15373.
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8

Multicolor Flow Cytometry Staining

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Allophycocyanin (APC)-conjugated and purified mouse anti-6×His (clone ad 1.1.10) was purchased from Abcam. Fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD38 (90/CD38), APC- and phycoerythrin (PE)-conjugated rat anti-mouse GL7 (GL7), PE-Cy7- and Pacific blue-conjugated rat anti-mouse CD45R/B220 (RA3-6B2), and Pacific blue-conjugated hamster anti-mouse CD3ε (500A2) were purchased from BD Biosciences. APC-conjugated streptavidin was purchased from eBioscience. DyLight 6470-conjugated AffiniPure goat anti-mouse IgG (all subclasses), Fcγ fragment specific, was purchased from Jackson ImmunoResearch.
Frank G.M., Angeletti D., Ince W.L., Gibbs J.S., Khurana S., Wheatley A.K., Max E.E., McDermott A.B., Golding H., Stevens J., Bennink J.R, & Yewdell J.W. (2015). A Simple Flow-Cytometric Method Measuring B Cell Surface Immunoglobulin Avidity Enables Characterization of Affinity Maturation to Influenza A Virus. mBio, 6(4), e01156-15.
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9

Comprehensive Immune Phenotyping Panel

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The following fluorescent conjugated anti-human monoclonal antibodies (mAbs) were utilized in this study: anti-CD3 (clone UCHT-1 and clone SK7), anti-CD56 (clone HCD56), anti-CD69 (clone FN50), anti-IFN-γ (clone 4S.B3) and anti-TNF-α (clone MAb11) mAbs were from Biolegend. Anti-NKp30 (clone Z25), anti-NKp44 (clone Z231), anti-NKG2A (clone Z199), anti-KIR2DL1/S1 (clone EB6B) and anti-KIR3DL1/S1 (clone Z27) mAbs were from Beckman Coulter. Anti-NKp46 (clone 9E2/NKp46), anti-KIR2DL2/L3/S2 (clone DX27), and anti-CD107a (clone H4A3) mAbs were from BD Bioscience. Anti-NKG2C (clone 134,591) mAb was from R&D system. Biotinylated anti-NKG2D (clone 1D11) mAb and APC conjugated streptavidin were from eBioscience.
Taechasan N., Scherwitzl I., Supasa P., Dejnirattisai W., Sriruksa K., Limpitikul W., Malasit P., Screaton G.R., Mongkolsapaya J, & Duangchinda T. (2024). The alteration of NK cells phenotypes related to the functions and dengue disease outcomes. Virus Research, 345, 199382.
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10

Bone Marrow Cell Biotin Retention Analysis

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Bone marrow cells were analyzed for retention of biotin labeling as previously described [26 (link)]. Cell surface markers used for analysis were identical to those used for cell cycle analysis with the following exceptions: biotinylated Ryk was replaced with sheep anti-mouse Ryk (AF4649, R&D Systems) with Alexa-Fluor 546-conugated donkey anti-sheep IgG acting as a secondary antibody (Invitrogen, Carlsbad, CA, http://www.invitrogen.com). APC-conjugated streptavidin (eBioscience) was used to detect biotin-labeled cells.
Povinelli B.J, & Nemeth M.J. (2014). Wnt5a Regulates Hematopoietic Stem Cell Proliferation and Repopulation Through the Ryk Receptor. Stem cells (Dayton, Ohio), 32(1), 105-115.
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