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Streptavidin pe cy5

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Streptavidin-PE-CY5.5 is a fluorescent dye-labeled streptavidin conjugate. Streptavidin binds to biotin with high affinity. The PE-CY5.5 fluorescent dye is attached to the streptavidin protein.

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Lab products found in correlation

Cal lyse, by Thermo Fisher Scientific (3 mentions) Golgistop, by BD (3 mentions) Brdu flow kit, by BD (2 mentions) Foxp3 pe, by Thermo Fisher Scientific (2 mentions) Ly6g apc cy7, by BioLegend (2 mentions) Anti mouse ly6c apc, by BioLegend (2 mentions) Cd11b biotin, by Thermo Fisher Scientific (2 mentions) Il 17 alexa fluor 647, by Thermo Fisher Scientific (2 mentions) Wt c57bl 6 mice, by Jackson ImmunoResearch (1 mentions) Anti cd4 apc efluor 780, by Thermo Fisher Scientific (1 mentions) Anti cd11c pe, by BioLegend (1 mentions) Anti cd5 apc, by BD (1 mentions) Anti tcrβ pe, by Thermo Fisher Scientific (1 mentions) Anti cd19 alexa 700, by Thermo Fisher Scientific (1 mentions) Anti cd11b brilliant violet 605, by BioLegend (1 mentions) Anti igm apc cy7, by BioLegend (1 mentions) Anti igm fitc, by Southern Biotech (1 mentions) Sba clonotyping system b6 c57j hrp, by Southern Biotech (1 mentions) 4 hydroxy 3 nitrophenylacetyl np ficoll, by Biosearch Technologies (1 mentions) Anti cd244, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Streptavidin-PE (135 citations) Streptavidin-Cy5 (38 citations) PE-Cy5.5-conjugated streptavidin (3 citations) PE-Cy5.5-streptavidin conjugate (2 citations)

12 protocols using streptavidin pe cy5

1

Comprehensive Cytometric Analysis of NK Cell Degranulation

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For cytometric analyses, cells were blocked with anti-CD16/32 (15 min at RT) that was followed by a 30-min incubation on ice with the specific antibodies. All antibodies used are listed in the Supplementary Table S1. For biotinylated antibodies, an additional 15-min incubation on ice was done with streptavidin-PE-CY5.5 (Ebioscience). BrDU stains were done 12 h after the injection of 1 mg BrDU using the BrDU Flow Kit (BD Biosciences). We determined NK cell degranulation rates by co-incubating lymphoma cells and splenocytes at equal numbers (106 each in 24-well plates, 500 μl) in the presence of anti-CD107a (5 h, 6.25 μg/ml) and of Golgistop (BD Pharmingen, present during the last 4 h). Erythrocytes were lysed in PBMC samples after antibody staining using CAL-LYSE (Invitrogen). Data analysis was performed using FlowJo software.
Dubois S., Waldmann T.A, & Müller J.R. (2020). Engagement of lymphoma T cell receptors causes accelerated growth and the secretion of an NK cell-inhibitory factor. Cellular immunology, 357, 104213.
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2

Cd244 Knockout Mice for Immunological Studies

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C57BL/6 (WT) mice were purchased from The Jackson Laboratories (Bar Harbor, ME).
Cd244-/- mice on the C57BL/6 background [12 (link)], were kindly provided by Dr. Raymond Welsh (University of Massachusetts Medical Center, Worcester, MA). Mice were maintained in the Translational Biomedical Research Center of the Medical College of Wisconsin and both genders were used for experiments in an age (6–12 week) and gender-matched manner. anti-CD45R-PE-Texas Red, anti-CD45R-PE, anti-GL7-FITC and anti-CD5-APC were purchased from BD Biosciences (San Jose, CA). CD21-eFluor 450, anti-CD23-PE-Cy7, anti-CD93-biotin, anti-CD4-APC-eFluor 780, anti-CD8-PE-Cy7, anti-TCRβ-FITC, anti-TCRβ-PE, anti-CD11b-Alexa Fluor 488, anti-CD244, anti-Foxp3-PE, anti-CD19-Alexa 700, anti-CD4-PE, anti-IgG-FITC and Streptavidin PE-Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-CD11b-Brilliant Violet 605, anti-CD11c-PE, anti-NK1.1-APC, anti-IgM-APC Cy7, anti-IgD-Pacific Blue, anti-CD38-Alexa Fluor 647, anti-CD138-APC and rat anti-mouse IgG2a-biotin were purchased from Biolegend (San Diego, CA). Anti-IgM-FITC and the SBA Clonotyping System-B6/C57J-HRP were purchased from Southern Biotech (Birmingham, AL). 4-Hydroxy-3-nitrophenylacetyl (NP)-ficoll, NP-Chicken Gamma Globulin (CGG), NP(24)-PE and NP-BSA were purchased from Biosearch Technologies (Novato, CA).
Ray A., Yuan C.Y., Miller N.M., Mei H, & Dittel B.N. (2015). 2B4 Is Dispensable for T-Dependent B Cell Immune Responses, but Its Deficiency Leads to Enhanced T-Independent Responses Due to an Increase in Peritoneal Cavity B1b Cells. PLoS ONE, 10(8), e0137314.
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3

Characterizing Autoimmune Neuroinflammation

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Myelin oligodendrocyte glycoprotein 35–55 (MOG35–55) peptide (MEVGWYRSPFSRVVHLYRNGK) and myelin basic protein (MBP) Ac1–11 peptide (Ac-ASQKRPSQRSK) were generated by the Protein Core Laboratory of the Blood Research Institute, BloodCenter of Wisconsin. KYC was synthesized by Biomatik (Wilmington, DE). The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780 CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse IFN-γ-PE was purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). The SMI-32 antibody, which detects nonphosphorylated neurofilament-H was purchased from Covance (Emeryville, CA). Streptavidin Alexa 405 and goat anti-mouse Alex 456 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-MPO heavy chain was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX). Anti-rabbit IgG (H + L)-HRP was purchased from Jackson ImmunoResearch (West Grove, PA). Anti-β-actin-HRP was purchased from Sigma-Aldrich (St. Louis, MO). For all experiments, the mice were age (6–8 wk) and gender matched with both sexes being utilized.
Zhang H., Ray A., Miller N.M., Hartwig D., Pritchard KA J.r, & Dittel B.N. (2015). Inhibition of Myeloperoxidase at the Peak of Experimental Autoimmune Encephalomyelitis Restores Blood-Brain-Barrier Integrity and Ameliorates Disease Severity. Journal of neurochemistry, 136(4), 826-836.
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4

Murine Model of Experimental Autoimmune Encephalomyelitis

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Myelin oligodendrocyte glycoprotein 35-55 (MOG35-55) peptide (MEVGWYRSPFSRVVHLYRNGK) was generated by the Protein Core laboratory of the Blood Research Institute, BloodCenter of Wisconsin. The 2.4G2 hybridoma was obtained from the American Tissue Culture Collection. Anti-mouse CD4-APC-eFluor 780, CD25-Alexa Fluor 700, CD11b-PE, CD11b-biotin, IL-17-Alexa Fluor 647, CD11c-Biotin, B220-Biotin, CD8-Biotin, CD11b-biotin, Foxp3-PE and streptavidin-PE Cy5.5 were purchased from eBioscience (San Diego, CA). Anti-mouse B220-PE-Texas Red, IFN-γ-PE, anti-active caspase 3-FITC and anti-human Ki67-FITC were purchased from BD Biosciences (San Diego, CA). Anti-mouse Ly-6C-APC and Ly-6G-APC-Cy7 were purchased from Biolegend (San Diego, CA). Anti-mouse/rat Neuropilin-1-APC was obtained from R&D Systems (Minneapolis, MN). Monoclonal antibodies SMI-32 (anti-nonphosphorylated neurofilament-H) and SMI-99 (anti-myelin basic protein (MBP)) were purchased from Covance (Emeryville, CA). Strepavidin Alexa 405 and goat anti-mouse Alexa 546 (H + L) were purchased from Life Sciences Advanced Technologies (St. Petersburg, FL). Anti-Biotin microbeads were purchased from MiltenyiBiotec (Auburn, CA).
Ray A., Basu S., Miller N.M., Chan A.M, & Dittel B.N. (2014). An Increase in Tolerogenic Dendritic Cell and Natural T Regulatory Cell Numbers During EAE in Rras−/− Mice Results in Attenuated Disease. Journal of immunology (Baltimore, Md. : 1950), 192(11), 5109-5117.
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5

Isolation and Characterization of Stem Cells

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Goat anti-mouse SCF and Nanog antibodies and rabbit anti-mouse antibodies were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). Anti-mouse CD45, Sca-1, c-kit, CD34, FLK2-, CD3, B220, TER-119-, antibodies (rat anti mouse) to various stem cell markers CD105, CD29 and CD44, along with secondary antibody streptavidin PE/Cy5.5, and donkey anti-rat –APC were obtained from eBioscience (San Diego, CA, USA). Rat anti mouse CD105 was obtained from Santa Cruz Biotechnology (Dallas, TX, USA). Biotin conjugated lineage antibody cocktail was obtained from Miltenyi Biotec (Miltenyi Biotec Inc, Auburn, CA). Bone morphogenetic protein-2 (BMP-2) was obtained from R&D Systems (R&D Systems, Minneapolis, MN). PLLA was obtained from Medisorb 100L 1A (Lakeshore Biomaterials, AL, USA) with inherent viscosity of 1.9 dL/g.
Shen J., Nair A., Saxena R., Zhang C.C., Borrelli J J.r, & Tang L. (2014). Tissue Engineering Bone Using Autologous Progenitor Cells in the Peritoneum. PLoS ONE, 9(3), e93514.
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6

NKT Cell Phenotyping by Flow Cytometry

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For cytometric analyses, cells were blocked with anti-CD16/32 (15 min at RT) that was followed by a 30-min incubation on ice with the specific antibodies. To identify CD1d-restricted NKT cells, cells were stained with a specific tetramer for 30 min at room temperature prior to antibody staining. All antibodies and the tetramer used are listed in the Supplementary Table S1. For biotinylated antibodies, an additional 15-min incubation on ice was done with streptavidin-PE-CY5.5 (Ebioscience). Liver cells were isolated by tissue homogenization using a syringe plunger that was followed by digestion in liver digest medium (Thermo Fisher Scientific, 15 min at 37°C) and Percoll centrifugation (37.5%, GE Healthcare Life Sciences). We determined NK cell degranulation rates by co-incubating lymphoma cells and splenocytes at equal numbers (106 each in 24-well plates, 500 μl) in the presence of anti-CD107a (5 h, 6.25 μg/ml) and of Golgistop (BD Pharmingen, present during the last 4 hours). NKG2D neutralization was done with HMG2D (Bio X Cell, 10 μg/ml). MHC class I expression was mimicked by pre-exposing SJ3 lymphoma cells to QDM peptide (MBL, 10 μg/ml, 10 min at 37°C) prior to co-culture. Erythrocytes were lysed in PBMC samples after antibody staining using CAL-LYSE (Invitrogen). Data analysis was performed using FlowJo software.
Dubois S., Feigenbaum L., Waldmann T.A, & Müller J.R. (2020). NK cells prevent T cell lymphoma development in T cell receptor-transgenic mice. Cellular immunology, 352, 104081.
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7

Comprehensive Cytometric Analysis of NK Cell Degranulation

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For cytometric analyses, cells were blocked with anti-CD16/32 (15 min at RT) that was followed by a 30-min incubation on ice with the specific antibodies. All antibodies used are listed in the Supplementary Table S1. For biotinylated antibodies, an additional 15-min incubation on ice was done with streptavidin-PE-CY5.5 (Ebioscience). BrDU stains were done 12 h after the injection of 1 mg BrDU using the BrDU Flow Kit (BD Biosciences). We determined NK cell degranulation rates by co-incubating lymphoma cells and splenocytes at equal numbers (106 each in 24-well plates, 500 μl) in the presence of anti-CD107a (5 h, 6.25 μg/ml) and of Golgistop (BD Pharmingen, present during the last 4 h). Erythrocytes were lysed in PBMC samples after antibody staining using CAL-LYSE (Invitrogen). Data analysis was performed using FlowJo software.
Dubois S., Waldmann T.A, & Müller J.R. (2020). Engagement of lymphoma T cell receptors causes accelerated growth and the secretion of an NK cell-inhibitory factor. Cellular immunology, 357, 104213.
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8

Bone Marrow and Spleen Cell Isolation and Quantification

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Single-cell suspensions from bone marrow or spleen were prepared by passing the tissues through 70 μm cell strainers (BD Pharmingen, San Diego, CA) and red blood cells were lysed in RBC lysis buffer (150 mM NH4Cl, 10 mM KHCO3, 0.1 mM EDTA). One million cells from each sample were then labeled with the following, anti-CD41-FITC (BD Pharmingen) or anti-TER119-APC-Cy7 (BD Pharmingen) for 30 minutes on ice. Labeled cells were washed in autoMACS Rinsing Solution (Miltenyi Biotec, Aubum, CA) with the addition of 0.5% BSA, fixed with 1% paraformaldehyde in PBS overnight and then analyzed on a LSRII cytometer (BD Biosciences, San Jose, CA). For analysis of Lineage-Sca1+c-Kit+ (LSK) cell and myeloid progenitor cells, ten million bone marrow cells were lineage depleted by using the Lineage Cell Depletion Kit (Miltenyi Biotec) according to the manufacturer's protocol with the addition of the anti-IL-7R-biotin antibody (BD Pharmingen). After magnetic separation of the lineage cells through the MACS MS column, the lineage negative cells were incubated with streptavidin-PE-Cy5.5 (eBioscience, San Diego, CA), anti-Sca1-PE-Cy7 (BD Pharmingen), anti-c-Kit-APC (BD Pharmingen), anti-FcγR-PE (BD Pharmingen) and anti-CD34-FITC (BD Pharmingen) on ice for 30 minutes and analyzed on a LSRII cytometer after washing and fixation as described above.
Liu C., Yu T., Xing Z., Jiang X., Li Y., Pao A., Mu J., Wallace P.K., Stoica G., Bakin A.V, & Yu Y.E. (2017). Triplications of human chromosome 21 orthologous regions in mice result in expansion of megakaryocyte-erythroid progenitors and reduction of granulocyte-macrophage progenitors. Oncotarget, 9(4), 4773-4786.
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9

Multiparameter Flow Cytometry Analysis of Leukemia

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For evaluation of MLL-AF9-transduced leukemia development, either peripheral blood or BM cells were stained with anti-Mac-1-APC, anti-Gr-1-PE, anti-CD3-APC, anti-B220-PE or anti-c-Kit-PE monoclonal antibodies (eBioscience) for the analyses of the myeloid and lymphoid lineages, differentiation, and cell frequencies of Mac-1 + c-Kit + LICs. For examination of Lin À CD127 À Sca-1 À c-Kit + CD16/32 + CD34 + L-GMP cells, BM leukemia cells were stained with biotinylated anti-CD3, anti-CD8, anti-B220, anti-Gr-1, anti-Ter119 and anti-CD127 antibody followed by staining with streptavidin-PE/Cy5.5, anti-Sca-1-PE/CY7, anti-c-Kit-APC, anti-CD16/ 32-eFluo450 and anti-CD34-PE (eBioscience). To determine the ratios of SoNar fluorescence, cells were excited at 405 nm/488 nm and acquisited at 525 nm (for SoNar sensor), or excited at 405 nm/561 nm and acquisited at 525 nm/610 nm (for SoNar-mCherry sensor), respectively. The mean fluorescence intensities and ratio (405/561) histograms were analyzed for the calculation of SoNar's signal by using FlowJo software v10. For the evaluation of homing ability, CXCR4 antibody (eBioscience) was used. The cell cycle status of SoNar-high and low AML cells were measured by staining with Hoechst 33342/Pyronin Y as previously described (Zheng et al., 2011) .
Hao X., Gu H., Chen C., Huang D., Zhao Y., Xie L., Zou Y., Shu H.S., Zhang Y., He X., Lai X., Zhang X., Zhou B.O., Zhang C.C., Chen G.Q., Yu Z., Yang Y, & Zheng J. (2019). Metabolic Imaging Reveals a Unique Preference of Symmetric Cell Division and Homing of Leukemia-Initiating Cells in an Endosteal Niche. Cell metabolism, 29(4).
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10

Mouse Hematopoietic Stem Cell Immunophenotyping

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Allophycocyanin-CY7-conjugated anti-mouse c-Kit antibody (2B8); PE-CY7-conjugated anti-mouse FcεRIα antibody (MAR-1); biotin-labeled anti-lineage marker antibodies (anti-CD3 (clone name: CT-CD3), anti-CD4 (RM4–5), anti-CD8 (53–6.7), anti-B220 antibody (RA3–6B2), anti-Gr-1 (RB6–8C5), anti-CD19 antibody (1D3), anti-IL-7Rα antibody (A7R34) antibodies; Alexa Fluor 647-conjugated anti-CD34 (RAM34); Pacific Blue-conjugated anti-mouse Sca-1 (D7); PE/Cyanine7-conjugated anti-mouse; FcγRII/III antibodies; streptavidin-PE-Cy5 were all purchased from eBioscience (San Diego, CA) or Biolegend (San Diego, CA).
Li Y., Qi X., Zhao D., Urban J.F, & Huang H. (2022). IL-3 Expands Pre-Basophil and Mast Cell Progenitors by Upregulating the IL-3 Receptor Expression. Cellular immunology, 374, 104498.
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