The lectin Alexa-Fluor 647® Wheat Germ Agglutinin (WGA-647 – Invitrogen – W32466) was suspended in sterile distilled water to obtain a stock solution at 1 mg/mL. A stock solution of Bodipy-FL Vancomycin (Van-Bodipy, Invitrogen – V34850) was prepared in DMSO at 0.5 mg/mL. Aliquotes of both stock solutions were stored at -20°C. For Gram analysis, the LIVE BacLightTM Bacterial Gram Stain Kit (Invitrogen – L7005) was also used as recommended by the manufacturer. This kit is composed of Syto 9 and HI at 3.34 and 4.67 mM, respectively. For viability analysis, the Live/Dead® BacLightTM bacterial viability kit (Invitrogen – L34856) was used as recommended by the manufacturer. This kit is composed of Syto 9 (3.34 mM in DMSO) and propidium iodide (PI) (20 mM in DMSO), two nucleic acid stains that stain all bacteria and in preference bacteria with a damaged cell wall that are considered as “dead,” respectively (Berney et al., 2007 (link)).
Alexa fluor 647 wheat germ agglutinin
Manufactured by Thermo Fisher Scientific
Alexa-Fluor 647® Wheat Germ Agglutinin is a fluorescent-labeled lectin that binds to N-acetylglucosamine and sialic acid residues on cell surfaces. It can be used to label and visualize cell surface glycoconjugates in various applications.
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Lab products found in correlation
Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions)
Van bodipy, by Thermo Fisher Scientific (1 mentions)
Observer d1 microscope, by Zeiss (1 mentions)
Green fluorescent protein, by Abcam (1 mentions)
Active nonphosphorylated β catenin, by Cell Signaling Technology (1 mentions)
Lsm 510 duo microscope, by Zeiss (1 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (1 mentions)
Zen 2012, by Zeiss (1 mentions)
Fsc 22 frozen section media, by Leica (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Hoechst 33342, by Thermo Fisher Scientific (1 mentions)
Cryostat, by Leica (1 mentions)
2 protocols using alexa fluor 647 wheat germ agglutinin
1
Gram Staining and Viability Analysis of Bacteria
2
Immunohistochemical Analysis of Cardiac Markers
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Immunohistochemical staining was performed on heart sections (5 μm). Antigen retrieval was carried out in sodium citrate buffer (pH 6.0) at 92°C using a BP‐111 laboratory microwave (Microwave Research and Applications). Immunostaining was performed with primary antibodies for Rac1 (Santa Cruz Biotechnology), phosphohistone‐H3 (phospho S10) (Abcam), cleaved caspase‐3 (Cell Signaling Technology), active (nonphosphorylated) β‐catenin (Cell Signaling Technology), and green fluorescent protein (Abcam). All slides were imaged with a Zeiss Observer D1 microscope using AxioVision Rel 4.7 software. For phalloidin and wheat germ agglutinin staining, P0 heart samples were fixed in 4% paraformaldehyde, cryoprotected in 30% sucrose, embedded in FSC22 frozen section media (Leica), and sectioned with a Leica cryostat at 10‐μm thick onto glass slides. Slides were incubated with Alexa Fluor 488 phalloidin (Life Technologies), Alexa Fluor 647 wheat germ agglutinin (Invitrogen), and Hoechst 33342 (Invitrogen). Confocal images were obtained with a Zeiss LSM 510 Duo microscope using ZEN 2012 software (Zeiss).
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