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Dylight 594 conjugated tomato lectin

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DyLight 594 conjugated Tomato Lectin is a fluorescent dye-labeled lectin used for the detection and visualization of glycoprotein-containing structures. It binds to N-acetylglucosamine and sialic acid residues on cell surfaces and extracellular matrices.

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Lab products found in correlation

Lsm 710, by Zeiss (2 mentions) α sma, by Thermo Fisher Scientific (1 mentions) Sp8 2 photon microscope, by Leica (1 mentions) Hp deepsee laser, by Spectra-Physics (1 mentions) Matlab, by MathWorks (1 mentions)

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DyLight 594 (25 citations) Tomato lectin (21 citations) Lectins (7 citations) DyLight 594 lectin (5 citations)

4 protocols using dylight 594 conjugated tomato lectin

1

Immunohistochemical Analysis of Ocular RAS

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Slides containing 8-µm-thick sagittal sections of the eye and eyelid were rinsed in PBS and blocked with PBS solution containing 2% bovine serum albumin for 30 minutes. The tissue sections were then incubated with primary antibodies for alpha-smooth muscle actin (α-SMA) (1:100 dilution; Invitrogen, Carlsbad, CA, USA), angiotensinogen (1:50 dilution; R&D Systems, Minneapolis, MN, USA), ACE (1:25 dilution; R&D Systems), AT1 receptors (1:50 dilution; Novus Biologicals, Littleton, CO, USA), and DyLight 594 conjugated tomato lectin (1:200 dilution; Vector Laboratories, Burlingame, CA, USA) for 90 minutes. The slides were washed with PBS three times and incubated with Alexa Fluor 488 or Alexa Fluor 647 conjugated secondary antibody (1:500 dilution; Abcam, Cambridge, UK) for 60 minutes. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). The slides were imaged using a confocal microscope (Nikon, Melville, NY, USA). The number of nuclei showing α-SMA staining and the percent fraction of area showing angiotensinogen and ACE staining were quantified using Image J (National Institutes of Health, Bethesda, MD, USA).
Shamloo K., Weng J., Ross C., Lee J., Alfuraih S., Totonchy J, & Sharma A. (2021). Local Renin-Angiotensin System Activation and Myofibroblast Formation in Graft Versus Host Disease–Associated Conjunctival Fibrosis. Investigative Ophthalmology & Visual Science, 62(13), 10.
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2

Visualizing Mouse Lymphatic Dynamics

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The mouse hindlimb was treated with depilating cream and washed with PBS to remove hair. After removal of the superficial layer of the skin, lymphatic collectors were visualized via injection of 10 to 20 μl (s.c.) of DyLight 594–conjugated tomato lectin (Vector Laboratories) into the footpad. Images of popliteal lymphatics were collected using a customized Leica SP8 2-photon microscope equipped with a ×25/0.95 NA water-dipping objective. A Mai Tai HP DeepSee Laser (Spectra- Physics) tuned to 900 nm provided the excitation light. Fluorescence emission was separated by dichroic mirrors at 458, 495, and 560 nm (Semrock) and directed to external detectors. 2-Photon excitation produced a second harmonic signal from collagen. Vessel diameters were measured using Imaris (Bitplane) and Matlab (Mathworks). After the 594 nm channel was isolated in Imaris, a Matlab script was used to break the vessel image into 20 sections, and then the width was measured randomly within each section. The average value of all 20 measurements gave the average vessel diameter at each point in time. Graphs were generated from the average thickness at each point over the 1000 frames collected at 14 fps.
Davis M.J., Kim H.J., Zawieja S.D., Castorena-Gonzalez J.A., Gui P., Li M., Saunders B.T., Zinselmeyer B.H., Randolph G.J., Remedi M.S, & Nichols C.G. (2020). Kir6.1-dependent KATP channels in lymphatic smooth muscle and vessel dysfunction in mice with Kir6.1 gain-of-function. The Journal of physiology, 598(15), 3107-3127.
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3

In Vivo Vasculature Labeling

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Before imaging, animals were anaesthetized with ketamine/ xylazine mixture and placed in the supine position and a small incision was made between the neck and shoulder. The jugular vein was exposed for injection of 100 μl of Dylight 594 conjugated Tomato Lectin (1mg/ml) (Vector Laboratories, CA). The lectin was allowed to circulate for 8–10 min after which the animal was sacrificed.
Bentolila L.A., Prakash R., Mihic-Probst D., Wadehra M., Kleinman H.K., Carmichael T.S., Péault B., Barnhill R.L, & Lugassy C. (2016). Imaging of Angiotropism/Vascular Co-Option in a Murine Model of Brain Melanoma: Implications for Melanoma Progression along Extravascular Pathways. Scientific Reports, 6, 23834.
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4

Visualizing Vascular Networks in Germinal Centers

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Immunized mice were injected i.v. with 150μg Dylight 594 conjugated Tomato Lectin (Vector Labs). After 10-30m, the mice were euthanized and perfused with 30ml of 1% PFA followed by 30ml of PBS. Spleens frozen in OCT compound were cryo-sectioned (5μm), air dried and fixed in 1:1 mix of acetone/methanol at -20°C/10m. Slides were warmed to RT, Pap Pen applied and re-hydrated (0.1% Tween 20, 0.5% BSA in 1xPBS) for 20m, Fc blocked for 20m, primary stained for 3h, washed 3x and stained with αFITC Oregon Green (Invitrogen) for 1h, washed 3x and mounted. Images were acquired on a Nikon e80i microscope or a Zeiss LSM 710. ImageJ FIJI was used for analysis. Mask files were created for GCs (GL7+), follicles (B220+), and vasculature (tomato lectin+). Vasculature mask was expanded radially to 40μm and percent GC or follicle area covered was determined.
Abbott R.K., Thayer M., Labuda J., Silva M., Philbrook P., Cain D.W., Kojima H., Hatfield S., Sethumadhavan S., Ohta A., Reinherz E.L., Kelsoe G, & Sitkovsky M. (2016). Germinal center hypoxia potentiates immunoglobulin class switch recombination. Journal of immunology (Baltimore, Md. : 1950), 197(10), 4014-4020.
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