Dylight 488 streptavidin

Manufactured by BioLegend

Sourced in United States

DyLight 488 Streptavidin is a fluorescently labeled protein used for the detection and visualization of biotinylated molecules. It has an excitation wavelength of 493 nm and an emission wavelength of 518 nm, resulting in a green fluorescent signal. DyLight 488 Streptavidin binds to biotin with high affinity, enabling its use in a variety of applications, such as immunoassays and flow cytometry.

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Lab products found in correlation

Alexa fluor647 conjugated anti mouse cd45r, by BioLegend (3 mentions) Pannoramic scan instrument, by 3DHISTECH (3 mentions) Biotinylated pna, by Vector Laboratories (2 mentions) Pierce cell surface protein isolation kit, by Thermo Fisher Scientific (2 mentions) Alexa fluor 568 conjugated f ab 2 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Hoechst 33342, by Thermo Fisher Scientific (2 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions) Goat serum, by Merck Group (2 mentions) Axio imager m2, by Zeiss (1 mentions) Biotinylated peanut agglutinin pna, by Vector Laboratories (1 mentions) Cryostat, by Leica (1 mentions) Anti gm130, by Proteintech (1 mentions) Anti pdi, by Cell Signaling Technology (1 mentions) Alexa fluor 488 conjugated donkey anti mouse igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 546 conjugated donkey anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Anti β catenin, by Proteintech (1 mentions) Mouse anti isl1, by Developmental Studies Hybridoma Bank (1 mentions) Mouse monoclonal anti qh1, by Developmental Studies Hybridoma Bank (1 mentions) Streptavidin cy3, by BioLegend (1 mentions) Ab9465, by Abcam (1 mentions)

Spelling variants of this product

Dylight 488 (3 citations) DyLight 488-conjugated streptavidin (2 citations)

8 protocols using dylight 488 streptavidin

Immunohistochemical Analysis of Splenic Germinal Centers

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The spleen of immunized mice was separated on day 7 after the final immunization and embedded in optimal cutting temperature (OCT) compound (SAKURA). The tissues were frozen with liquid nitrogen before sectioning (7 µm) on a cryostat. After being fixed in cold acetone and blocked with 5% FBS in PBS at RT for 1 h, the sections were incubated with Biotinylated PNA (VECTOR, 1:100) overnight at 4°C. DyLight 488 Streptavidin (BioLegend, 1:100) was used as the secondary antibody at RT for 1 h followed with Alexa Fluor647-conjugated anti-mouse CD45R (BioLegend, 1:150) at RT for 1 h. After staining, the sections were scanned under a Pannoramic SCAN instrument (3DHISTECH, Hungary).

Gao F., Huang J., Li T., Hu C., Shen M., Mu S., Luo F., Song S., Hao Y., Wang W., Han X., Qian C., Wang Y., Wu R., Li L., Li S, & Jin A. (2021). A Highly Conserved Peptide Vaccine Candidate Activates Both Humoral and Cellular Immunity Against SARS-CoV-2 Variant Strains. Frontiers in Immunology, 12, 789905.

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Biotin-Functionalized Microparticles for Cell Targeting

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To assess the functionalization with thiol-PEG-biotin, the modified MeLam microparticles were incubated with DyLight 488 Streptavidin (BioLegend, USA) prepared at a concentration of 10μg/ml in PBS for 15 minutes. Subsequently the microparticles were washed with PBS and observed under fluorescence microscopy (Axio Imager M2, Zeiss, Germany). Polymeric microparticles with covalently attached biotin are herein proposed as versatile targeting vehicles for multiple biomolecules; in this work the microparticles were functionalized with the tripeptide Arg-Gly-Asp (RGD) to increase cell adhesion. Briefly, biotinylated microparticles were incubated with purified streptavidin (Promega, USA) (25μg/ml) in PBS under gently agitation for 15min at room temperature. A washing step was then performed to remove unbound streptavidin. Finally, the MeLam microparticles were incubated with biotinylated RGD (25 μg/mL) (AnaSpec, USA) in PBS under gently agitation for 15min at room temperature. The MeLam microparticles were finally washed with PBS to remove unbound biotinylated RGD.

Martins C., Custódio C, & Mano J. (2018). Multifunctional laminarin microparticles for cell adhesion and expansion. Carbohydrate polymers, 202, 91-98.

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Immunohistochemical Analysis of Murine B Cells

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The spleens of immunized mice were separated on day 7 after the final immunization and embedded in Optimal Cutting Temperature (O.C.T) compound (SAKURA: #4583). The tissues were frozen at liquid nitrogen before sectioning (7 um) on a cryostat. After being fixed in cold acetone and blocked with 5% FBS in PBS at room temperature (RT) for 1 hour, the sections were incubated with Biotinylated PNA (VECTOR: #FL-1071-5, 1: 100) overnight at 4°C. DyLight 488 Streptavidin (BioLegend: #405218, 1: 100) was used as the secondary antibody at RT for 1 hour followed with Alexa Fluor647-conjugated anti-mouse CD45R (BioLegend: #103226,1: 150) at RT for 1 hour. After staining, the sections were scanned under a Pannoramic SCAN instrument (3DHISTECH, Hungary).

Gao F.X., Wu R.X., Shen M.Y., Huang J.J., Li T.T., Hu C., Luo F.Y., Song S.Y., Mu S., Hao Y.N., Han X.J., Wang Y.M., Li L., Li S.L., Chen Q., Wang W, & Jin A.S. (2022). Extended SARS-CoV-2 RBD booster vaccination induces humoral and cellular immune tolerance in mice. iScience, 25(12), 105479.

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Immunohistochemical Analysis of Germinal Centers

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The spleens from BALB/c mice were separated one week after the final immunization and embedded in Optimal Cutting Temperature (OCT) Compound (SAKURA, USA). The tissues were frozen at -80°C before sectioning (8 μm) on a Cryostat (Leica, Germany). After being fixed in cold acetone and blocked with 1% BSA in PBS at room temperature for 1 h, the sections were incubated with Biotinylated Peanut Agglutinin (PNA, 1 : 100 dilution, VECTOR, USA) at 4°C overnight. DyLight™ 488 Streptavidin (1 : 100 dilution, BioLegend, USA) was used as the secondary antibody at room temperature for 1 h. At last, Alexa Fluor® 647-conjugated anti-mouse CD45R (1 : 150 dilution, Clone RA3-6B2, BioLegend, USA) was incubated at room temperature for 1 h. The sections were scanned under a Pannoramic SCAN instrument (3DHISTECH, Hungary). For quantification the area of GCs, spleen sections of three mice from each group were analyzed by ImageJ software.

Gu Y., Sun X., Huang J., Zhan B, & Zhu X. (2020). A Multiple Antigen Peptide Vaccine Containing CD4+ T Cell Epitopes Enhances Humoral Immunity against Trichinella spiralis Infection in Mice. Journal of Immunology Research, 2020, 2074803.

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Immunofluorescence Staining of Cell Markers

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After fixing in 4% paraformaldehyde for 10 min, cells were treated with 0.2% Triton X-100 for 10 min at room temperature, followed by blocking with goat serum at 37 °C for 60 min. The cells were then immunostained with the appropriate antibody or lectin. The following primary antibodies or lectins were used: anti-GALNT1 (1:200; Abnova, CAT#H00002589-M10), anti-β-catenin (1:200; Proteintech, CAT#51067), anti-GM130 (1:200; Proteintech, CAT#11308-1-AP-150), anti-PDI (1:200; Cell Signaling Technology, CAT# 3501T), and VVA (1:200; Vector Laboratories, CAT# B-1235-2). Alexa Fluor 546-conjugated donkey anti‐rabbit IgG (1:1000, Thermo Fisher, CAT#A11003), DyLight™ 488 streptavidin (1:200, BioLegend, CAT# 405218), and Alexa Fluor 488-conjugated donkey anti-mouse IgG (1:1000, Thermo Fisher, CAT#A11001) were used as secondary antibodies. DAPI nucleic acid stain (Abcam, CAT# ab104139) was used to counterstain the sections. An inverted microscope was used to capture the images.

Zhang J., Wang H., Wu J., Yuan C., Chen S., Liu S., Huo M., Zhang C, & He Y. (2022). GALNT1 Enhances Malignant Phenotype of Gastric Cancer via Modulating CD44 Glycosylation to Activate the Wnt/β-catenin Signaling Pathway. International Journal of Biological Sciences, 18(16), 6068-6083.

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Cell Surface Protein Biotinylation Assay

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Biotinylation was performed using a Pierce Cell Surface Protein Isolation Kit (Thermo Fisher Scientific). MEF cells plated on 35 mm glass-bottom plates (MatTEK) were washed twice with ice cold PBS²⁺/BSA (PBS supplemented with 1.5 mM MgCl₂, 0.2 mM CaCl2, 0.18% (w/v) glucose, 0.2% (w/v) BSA). EZ-Link Sulfo-NHS-SS-Biotin was diluted in PBS²⁺/BSA (0.25 mg/ml) and added onto MEF cells for 30 min on a rotator in 4°C. Quenching solution was added (50 μl/ml biotin solution) to the cells and incubated for 15 min. The cells culture medium was changed with PBS²⁺/BSA and incubated in 37°C for 3 hours. Cells were washed with PBS followed by fixation with 4% (w/v) paraformaldehyde (Electron Microscopy Sciences). The samples were blocked with 10% (v/v) goat serum (MilliporeSigma) for 10 min then with anti Na⁺/K⁺-ATPase-α1 primary antibody at 4 °C overnight. The samples were washed multiple times with PBS for 3 hours at room temperature then incubated with the Alexa Fluor 568 conjugated F(ab℉)2-Goat anti-Mouse IgG (Life Technologies) for 1 hour at room temperature. The samples were washed with PBS for 3 hours at room temperature, incubated with DyLight™ 488 Streptavidin (BioLegend) for 30 min at room temperature and washed with PBS for 1hour at room temperature. Nuclei were stained with Hoechst 33342 (Thermo Fisher Scientific) for 10 min.

Chin A.C., Gao Z., Riley A.M., Furkert D., Wittwer C., Dutta A., Rojas T., Semenza E.R., Felder R.A., Pluznick J.L., Jessen H.J., Fiedler D., Potter B.V., Snyder S.H, & Fu C. (2020). The inositol pyrophosphate 5-InsP7 drives sodium-potassium pump degradation by relieving an autoinhibitory domain of PI3K p85α. Science Advances, 6(44), eabb8542.

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Immunostaining of Quail Embryo Sections

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Sections (14 μm) were prepared from frozen embryos, and were immunostained using the following antibodies: mouse monoclonal anti-quail cell (QCPN; Developmental Studies Hybridoma Bank; 1:100), mouse monoclonal anti-QH1 (Developmental Studies Hybridoma Bank; 1:100), mouse anti-Isl1 (Developmental Studies Hybridoma Bank; 1:100), mouse anti-MHC (MF20; Developmental Studies Hybridoma Bank; 1:100), mouse anti-sarcomeric α actinin (ab9465; Abcam; 1:100), rabbit anti-MHC (ab124205; Abcam; 1:100), rabbit anti-cardiac troponin I antibody (ab47003; Abcam; 1:100), rabbit anti-Nkx2.5 (ab35842; Abcam; 1:100). Signals were visualized with FITC (fluorescein isothiocyanate)- or TRITC-conjugated secondary antibodies specific for the appropriate species. Some sections were treated with biotin-conjugated secondary antibodies and visualized using the Cy3 streptavidin (Biolegend; 1:1000) or Dylight 488 streptavidin (Biolegend; 1:500). Nuclei were visualized with TO-PRO-3 Iodide (Molecular Probes; 1:1000). Fluorescent signals were visualized with a computer-assisted confocal microscope (Nikon ECLIPSE C2/Ti) and software (Nikon NIS-Elements AR4.100).

Asai R., Haneda Y., Seya D., Arima Y., Fukuda K., Kurihara Y., Miyagawa-Tomita S, & Kurihara H. (2017). Amniogenic somatopleure: a novel origin of multiple cell lineages contributing to the cardiovascular system. Scientific Reports, 7, 8955.

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Cell Surface Protein Biotinylation and Imaging

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Biotinylation was performed using the Pierce Cell Surface Protein Isolation Kit (Thermo Fisher Scientific). MEF cells plated on 35-mm glass-bottom plates (MatTEK) were washed twice with ice-cold PBS²⁺/BSA [PBS supplemented with 1.5 mM MgCl₂, 0.2 mM CaCl₂, 0.18% (w/v) glucose, and 0.2% (w/v) BSA]. EZ-Link Sulfo-NHS-SS-Biotin was diluted in PBS²⁺/BSA (0.25 mg/ml) and added onto MEF cells for 30 min on a rotator at 4°C. Quenching solution was added [biotin solution (50 μl/ml)] to the cells and incubated for 15 min. The cells culture medium was changed with PBS²⁺/BSA and incubated at 37°C for 3 hours. Cells were washed with PBS followed by fixation with 4% (w/v) paraformaldehyde (Electron Microscopy Sciences). The samples were blocked with 10% (v/v) goat serum (MilliporeSigma) for 10 min and then with anti–Na⁺/K⁺-ATPase-α1 primary antibody at 4°C overnight. The samples were washed multiple times with PBS for 3 hours at room temperature and then incubated with the Alexa Fluor 568–conjugated F(ab′)₂-Goat anti-Mouse IgG (Life Technologies) for 1 hour at room temperature. The samples were washed with PBS for 3 hours at room temperature, incubated with DyLight 488 Streptavidin (BioLegend) for 30 min at room temperature, and washed with PBS for 1 hour at room temperature. Nuclei were stained with Hoechst 33342 (Thermo Fisher Scientific) for 10 min.

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