Dylight 800

Manufactured by Thermo Fisher Scientific

Sourced in United States

DyLight 800 is a near-infrared fluorescent dye produced by Thermo Fisher Scientific. It is designed for use in a variety of biological and analytical applications, including protein labeling, immunoassays, and in vivo imaging. The dye has an excitation maximum at 777 nm and an emission maximum at 794 nm, making it suitable for detection in the near-infrared region of the spectrum.

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Lab products found in correlation

57 protocols using dylight 800

Fluorescent Secondary Antibody Dilutions

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1:10,000 goat anti-rabbit IgG (H+L) Dylight 800 (ThermoFisher, A32735), 1:10,000 goat anti-mouse IgG (H+L) Dylight 800 (ThermoFisher, A32730), 1:10,000 goat anti-rabbit IgG (H+L) Dylight 680 (ThermoFisher, A32734), 1:10,000 goat anti-mouse IgG (H+L) Dylight 680 (ThermoFisher, A32729), 1:10,000 goat anti-rat IgG (H+L) Dylight 800 (ThermoFisher, sa510024).

Llobet Rosell A., Paglione M., Gilley J., Kocia M., Perillo G., Gasparrini M., Cialabrini L., Raffaelli N., Angeletti C., Orsomando G., Wu P.H., Coleman M.P., Loreto A, & Neukomm L.J. (2022). The NAD+ precursor NMN activates dSarm to trigger axon degeneration in Drosophila. eLife, 11, e80245.

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Antibody Staining Protocol for Cell Analysis

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The following primary antibodies were used in this study: CC2C6 (Biolegend), B6H12 (BD Pharmingen) and BRIC126 (Santa Cruz Biotechnology) are α-CD47 monoclonals; Mcl-1, PARP-1 (Santa Cruz Biotechnology); NOXA (Cell Signaling); 9F10 (ITGA4), P84 (SIRPα), GAPDH, and Alexa Fluor 647-labelled F4/80 (Biolegend). Secondary antibodies used were Dylight488, Dylight633, Dylight680 and DyLight800 conjugates of goat-anti-mouse, and DyLight800 conjugated goat-anti-rabbit (all from Thermo Fisher).

Leclair P., Liu C.C., Monajemi M., Reid G.S., Sly L.M, & Lim C.J. (2018). CD47-ligation induced cell death in T-acute lymphoblastic leukemia. Cell Death & Disease, 9(5), 544.

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Serological Detection of T. gondii

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Serum from blood samples taken from each mouse were stored at −80 °C until tested for the presence of anti-T. gondii antibodies. Protein lysate from 5 × 10⁶ parasites of each strain were run on standard 10% acrylamide SDS-PAGE gels and transferred to PVDF membrane. The PVDF membranes were then cut into strips, blocked with 5% powdered milk, and each strip individually incubated with serum from each infected mouse at a dilution of 1:50. After washing, all strips were incubated in donkey anti-mouse secondary antibody conjugated with DyLight 800 (Life Technologies), washed, and then imaged on an Odyssey CLx (Li-COR).

McGovern K.E., Cabral C.M., Morrison H.W, & Koshy A.A. (2020). Aging with Toxoplasma gondii results in pathogen clearance, resolution of inflammation, and minimal consequences to learning and memory. Scientific Reports, 10, 7979.

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Fluorescent Immunohistochemistry Labeling Protocol

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The IL13Rα2 Ab was generated by Pfizer Inc. The fluorophores VivoTag680XL^® (VT680) (#NEV11119) and VivoTag645^® (VT645) (#NEV11173) were obtained from Perkin Elmer. AlexaFluor647^® (AF647) (#A-20006), AlexaFluor680^® (AF680) (#A-20008), AlexaFluor750^® (AF750) (#A-20011), BODIPY-X630/650^® (BOD630) (#D-10000), DyLight800^® (#46421), rhodamine-conjugated phalloidin (#R415) and Prolong antifade mounting media (#P36962) were obtained from Life Technologies, and IRDye^®800 (IRDye800) (#929-70020) was obtained from Licor. The Nunc^™ Lab-Tek^™ II Chamber Slide^™ System (#12-565-8), DAPI and an anti-human fluorescein-conjugated secondary Ab against human IgG (#PI31529) were obtained from Thermo Fisher Scientific Inc. BIOT sheep anti-human IgG polyclonal Ab (#AU003.MCUS01) and AlexaFluor647^® goat anti-human IgG (#A80-319A) were obtained from Binding Site and Bethyl Labs, respectively.

Gupta P., Wentland J.A., Leal M., Ma D., Roach R., Esparza A., King L., Spilker M.E., Bagi C., Winkelmann C.T, & Giddabasappa A. (2017). Assessment of near-infrared fluorophores to study the biodistribution and tumor targeting of an IL13 receptor α2 antibody by fluorescence molecular tomography. Oncotarget, 8(34), 57231-57245.

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Monitoring Ipa Protein Secretion

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The secretion of Ipa proteins after Congo red induction was carried our as previously described [11 (link)]. The total level of protein expression was revealed via western blot using monoclonal anti-IpaB H16 [57 (link)] and polyclonal anti-IpaH [58 (link)], anti-RNAP α subunit (gift from Prof. Akira Ishihama, Hosei University, Koganai, Tokyo) and anti-λ cI (gift from Prof. Ann Hochschild, Harvard Medical School, Boston). Goat anti-mouse DyLight 800 (Fisher Scientific) or goat anti-rabbit Alexa 680 (Invitrogen) conjugates were used as secondary antibodies. The membranes were then visualized using an Odyssey infrared imaging system (LI-COR Biosciences).

Shen D.K, & Blocker A.J. (2016). MxiA, MxiC and IpaD Regulate Substrate Selection and Secretion Mode in the T3SS of Shigella flexneri. PLoS ONE, 11(5), e0155141.

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SDS-PAGE and Western Blot Analysis

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SDS–PAGE and Western blot analysis were performed exactly as described previously (Barbosa et al., 2015 (link)). Antibodies used were as follows. Primary antibodies were mouse anti-V5, (1:10,000; Life Technologies), rabbit anti–actin 20-36 (1:2500; Sigma-Aldrich), mouse anti–actin AC-40 (1:2500; Sigma-Aldrich), rabbit anti–γ-tubulin (1:2500; Sigma-Aldrich), anti–ApoA-IV 1D6B6 (1:2000; Cell Signaling, Leiden, Netherlands), rabbit phospho-GSK-3α/β(Ser-21/9) D17D2 (1:2000; Cell Signaling), and anti–CREB-H pS73 and pS81 (1:100; Barbosa et al., 2015 (link)). Secondary antibodies were conjugated with horseradish peroxidase (for detection by conventional chemiluminescence) or with Dylight680 or Dylight800 (Fisher Scientific, Lutterworth, United Kingdom) for detection by laser scanning using the LI-COR Odyssey Image system. The latter method was routinely employed for CREB-H detection using the phosphospecific antibodies because it allowed simultaneous analysis, on one blot, of total CREB-H species with the anti-mouse antibody to the epitope tag in one channel and the phosphorylated species with the anti-rabbit antibody to the phosphorylated peptide in a second channel.

Barbosa S., Carreira S, & O’Hare P. (2017). GSK-3–mediated phosphorylation couples ER–Golgi transport and nuclear stabilization of the CREB-H transcription factor to mediate apolipoprotein secretion. Molecular Biology of the Cell, 28(11), 1565-1579.

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VEGFR2 Phosphorylation Assay in Cells

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HPAEC or mouse brain microvascular cell lysates were prepared in MgALB150 lysis buffer (50 mmol/L Tris pH 7.4, 150 mmol/L NaCl, 0.5% NP‐40, 5 mmol/L MgCl₂, phosphatase and protease inhibitors). Protein quantification was performed using a bicinchoninic acid assay kit (Thermo Fisher), and equivalent amounts of total cell protein were added to tubes containing 2 μg monoclonal rabbit anti‐VEGFR2 and 20 μL of protein G‐Sepharose beads. After incubation at 4°C overnight, immunoprecipitated proteins were blotted against phospho‐tyrosine (Cell Signaling) and then re‐probed with anti‐VEGFR2 to assess precipitation efficiency. Alternatively, lysates were directly probed with rabbit anti‐phosphoY1175 VEGFR2 (Thermo Fisher Scientific), followed by incubation of secondary antibody (goat anti‐rabbit DyLight‐800 (Fisher)). An Odyssey Infrared Imaging System (LI‐COR Biosciences) was used to image membranes and for densitometry.

DiStefano P.V, & Glading A.J. (2019). VEGF signalling enhances lesion burden in KRIT1 deficient mice. Journal of Cellular and Molecular Medicine, 24(1), 632-639.

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Monitoring Ipa Protein Secretion

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The secretion of Ipa proteins after Congo red induction was carried our as previously described37 (link). The total level of protein expression was revealed via western blot using either monoclonal anti-FLAG (Sigma) or polyclonal anti-MxiN23 (link). Goat anti-mouse DyLight 800 (Fisher Scientific) or goat anti-rabbit Alexa 680 (Invitrogen) conjugates were used as secondary antibodies. The membranes were then visualized using an Odyssey infrared imaging system (LI-COR Biosciences).

Makino F., Shen D., Kajimura N., Kawamoto A., Pissaridou P., Oswin H., Pain M., Murillo I., Namba K, & Blocker A.J. (2016). The Architecture of the Cytoplasmic Region of Type III Secretion Systems. Scientific Reports, 6, 33341.

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Quantitative Immunoblot Analysis of Signaling Proteins

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Gel electrophoresis and Western blotting were performed as previously described (46 ). In brief, after transfer, the membrane was incubated with Tris-buffered saline buffer supplemented with 0.1% Tween-20 (TBS-T) and 5% BSA at room temperature for 30 min on a shaker, then probed with primary antibodies (anti-PLCβ4 [Sigma–Aldrich; catalog no.: HPA007951], anti-Gαs [Sigma–Aldrich; catalog no.: 06-237], anti-Gβ [in house], anti-GAPDH [Invitrogen; catalog no.: MA5-15738]) diluted 1:1000 in TBS buffer supplemented with 0.1% Tween-20 and 3% BSA at 4 °C overnight. The membrane was washed with TBS-T four times and probed with secondary antibody (goat anti-rabbit immunoglobulin G, DyLight 800 (Invitrogen; catalog no.: SA535571) at room temperature for 1 h. After another four washes with TBS-T, immunoreactive proteins were visualized using Li-Cor Odyssey CLx and analyzed using Image Studio Lite software (Li-Cor).

Phan H.T., Loomis J., Abraham S., He Q., Bastepe M, & Smrcka A.V. (2022). A naturally occurring membrane-anchored Gαs variant, XLαs, activates phospholipase Cβ4. The Journal of Biological Chemistry, 298(8), 102134.

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Western Blotting of LysA Protein

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Western blotting assay was carried out essentially as described previously (23 (link)). Anti-gp10 antibodies were used as primary antibodies for probing LysA protein and its variant. Goat anti-Rabbit IgG (H+L) Secondary Antibody, DyLight 800 (catalog no. SA5-35571; Invitrogen) was used as secondary antibody. The blot was imaged on LI-COR image scanner.

Nair G, & Jain V. (2024). An intramolecular cross-talk in D29 mycobacteriophage endolysin governs the lytic cycle and phage-host population dynamics. Science Advances, 10(6), eadh9812.

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