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Fluorescein di β d galactopyranoside

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Fluorescein di-β-D-galactopyranoside is a chemical compound used as a fluorescent substrate in various biological and biochemical applications. It is composed of a fluorescein molecule attached to two galactose moieties. This compound can be used to detect the presence and activity of enzymes, such as β-galactosidase, which cleave the galactose groups and release the fluorescent fluorescein.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions) Bafilomycin a1, by Merck Group (2 mentions) Senescence β galactosidase staining kit, by Cell Signaling Technology (1 mentions) Lsrfortessa flow cytometer, by BD (1 mentions) Stain buffer, by BD (1 mentions) Anti cd45, by BD (1 mentions) Anti cd3, by BD (1 mentions) Anti cd8a, by BD (1 mentions) Anti b220, by BD (1 mentions) Anti cd4, by BD (1 mentions) Anti cd69, by BD (1 mentions) Anti f4 80, by BD (1 mentions) Anti cd11b, by BD (1 mentions) Anti ly6g, by BD (1 mentions) Anti ly6c, by BD (1 mentions) Tween 20, by Merck Group (1 mentions) Superblock buffer, by Thermo Fisher Scientific (1 mentions) Tmb substrate, by Thermo Fisher Scientific (1 mentions) Teflon af, by DuPont (1 mentions)

Close spelling variants of this product

Fluorescein di-β-D-galactopyranoside (FDG) (10 citations) D-fluorescein (2 citations)

5 protocols using fluorescein di β d galactopyranoside

1

Histological Analysis of Cardiomyocytes

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For histological analysis, embryos or heart tissues were fixed in 10% paraformaldehyde (PFA) at the stages indicated in each figure legend, dehydrated and embedded in paraffin for preparation of 5- or 10-μm histological sections. Rehydrated slides were stained with haematoxylin and eosin and Masson’s trichrome. For determination of CM cross-sectional area, deparaffinized and rehydrated heart sections were incubated for 1 h at room temperature with fluorescein isothiocyanate (FITC)-labelled wheatgerm agglutinin (Sigma-Aldrich) to visualize myocyte membranes. Regions that included the circular shapes of capillaries were selected from the epicardial side of the LV-free walls. The mean cross-sectional area of CMs was determined from 60 to 80 cells. For fluorescence detection of SA-β-gal activity, cells
(107 cell ml−1) were incubated with C12FDG (fluorescein di-β-D-galactopyranoside; 33 μM; Sigma), a β-galactosidase substrate that generates a fluorescent product upon cleavage, for 60 min at 37 °C (ref. 35 (link)). Cytochemical detection of senescent cells in vitro was determined in cells and fixed tissues with the Senescence β-Galactosidase Staining Kit (Cell Signaling).
Gonzalez-Valdes I., Hidalgo I., Bujarrabal A., Lara-Pezzi E., Padron-Barthe L., Garcia-Pavia P., Gómez-del Arco P., Redondo J.M., Ruiz-Cabello J.M., Jimenez-Borreguero L.J., Enriquez J.A., de la Pompa J.L., Hidalgo A, & Gonzalez S. (2015). Bmi1 limits dilated cardiomyopathy and heart failure by inhibiting cardiac senescence. Nature Communications, 6, 6473.
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2

Lysosomal Alkalization and β-Galactosidase Assay

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Cells were incubated in culture medium containing 100 nM Bafilomycin A1 (B1793, Sigma‐Aldrich) for 90 min to induce lysosomal alkalization. The fluorogenic substrate for β‐galactosidase, fluorescein di‐β‐D‐galactopyranoside (33 μM; F2756, Sigma‐Aldrich) or DDAO galactoside (10 μM; Setareh Biotech LLC), was subsequently added to the medium for 90 min. Cells were fixed in 4% paraformaldehyde for 15 min, rinsed with PBS, and permeabilized with 0.1% Triton X‐100 in PBS for 15 min. 0.5 μg/ml of DAPI (Sigma‐Aldrich) was used to counterstain DNA, and coverslips were then mounted on slides.
Barroso‐Vilares M., Macedo J.C., Reis M., Warren J.D., Compton D, & Logarinho E. (2020). Small‐molecule inhibition of aging‐associated chromosomal instability delays cellular senescence. EMBO Reports, 21(5), e49248.
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3

Multi-parameter Flow Cytometric Analysis of Immune Cell Populations

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Single-cell suspensions were resuspended in BD stain buffer (BD Bioscience) for 15 min prior to staining with specific antibodies. Antibodies against cell markers: anti-CD45, anti-CD3, anti-CD4, anti-CD8a, anti-CD11b, anti-F4/80, anti-Ly6G, anti-Ly6C, anti-NKp46(CD335), anti-B220, and anti-CD69, were purchased from BD Bioscience and BioLegend; anti-OX40L and anti-41BBL from Miltenyi Biotec (detailed in Supplementary table 4). Samples were mixed with FVS700 (1/7000) and data were acquired on an LSR Fortessa flow cytometer (BD Biosciences) and analyzed with FlowJo software (Tree Star). ROS generation in cultured neutrophils was determined using either DCFDA/H2DCFDA-Cellular ROS Assay Kit (ab113851) following the manufacturer’s protocol or a dihydroethidium (DHE) fluorescent probe. Cells were incubated with DHE (10 μM) in HBSS containing 1.5 mM CaCl2 and 1 mM MgCl2 for 30 min at 37 °C and analyzed by flow cytometry. LacZ activity was determined by flow cytometry using Fluorescein di[β-D-galactopyranoside (Sigma, F2756) as described elsewhere94 (link).
Uyanik B., Goloudina A.R., Akbarali A., Grigorash B.B., Petukhov A.V., Singhal S., Eruslanov E., Chaloyard J., Lagorgette L., Hadi T., Baidyuk E.V., Sakai H., Tessarollo L., Ryffel B., Mazur S.J., Lirussi F., Garrido C., Appella E, & Demidov O.N. (2021). Inhibition of the DNA damage response phosphatase PPM1D reprograms neutrophils to enhance anti-tumor immune responses. Nature Communications, 12, 3622.
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4

Quantifying Lysosomal β-Galactosidase Activity

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In fixed-cell analysis, cells were incubated for 90 min in medium with 100 nM Bafilomycin A1 (B1793, Sigma-Aldrich, MO, USA) to induce lysosomal alkalinization. Fluorogenic substrate (33 μM) for β-galactosidase, fluorescein di-β-d-galactopyranoside (F2756, Sigma-Aldrich, MO, USA) were then added to the medium, and incubation carried out for 90 min. Cells were fixed in 4% paraformaldehyde for 15 min, rinsed with PBS, and permeabilized with 0.1% Triton-X100 in PBS for 15 min. Finally, cells were counterstained with 1 µg/ml DAPI (Sigma-Aldrich, MO, USA). For FACS, cells were incubated in Bafilomycin A1 as described above and then exposed to 10 μM of fluorogenic substrate for β-galactosidase, DDAOG (9H-(1,3-dichloro-9,9-dimethylacridin-2-one-7-yl) β-d-Galactopyranoside), for 90 min (Setareh Biotech LLC, USA).
Macedo J.C., Vaz S., Bakker B., Ribeiro R., Bakker P.L., Escandell J.M., Ferreira M.G., Medema R., Foijer F, & Logarinho E. (2018). FoxM1 repression during human aging leads to mitotic decline and aneuploidy-driven full senescence. Nature Communications, 9, 2834.
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5

Integrated Biosensing Platform Fabrication

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Following reagents were purchased from Sigma-Aldrich (Belgium): all buffer components, bovine serum albumin (BSA), Tween 20, Krytox 1525 oil and Fluorescein-di-(β-D-galactopyranoside) (FDG). Teflon-AF was obtained from Dupont (United States). Parylene-C dimer and Silane A174 were purchased from Plasma Parylene Coating Services (Germany). Reagents for photolithography were obtained from Rohm and Haas (United States). The fluoroalkylsilane, Dynasylan F8263, was a kind gift from Evonik Degussa International AG (Essen, Germany). Dynabeads M280 Tosylactivated superparamagnetic beads were purchased from Life Technologies (United States). Superblock buffer, TMB substrate and β-galactosidase labeled streptavidin (SBG) were purchased from Thermo Scientific (United States). PlusOne Drystrip Coverfluid oil was obtained from GE Healthcare (The Netherlands). Abbott Architect TSH reagent kits were kindly provided by Abbott Laboratories (United States). Commercially available anti-TSH antibodies 5409 and T25C were purchased from Medix Biochemica (Finland) and Fitzgerald Industries International (United States), respectively. Magnets for the off chip experiments were obtained from Supermagnete (Webcraft GmbH, Germany). Cyanoethyl pullulan (CEP) was obtained from Shin-Etsu Chemical Co. Fluoropel PFC 1101V was purchased from Cytonix.
Leirs K., Dal Dosso F., Perez-Ruiz E., Decrop D., Cops R., Huff J., Hayden M., Collier N., Yu K.X.Z., Brown S, & Lammertyn J. (2022). Bridging the Gap between Digital Assays and Point-of-Care Testing: Automated, Low Cost, and Ultrasensitive Detection of Thyroid Stimulating Hormone. Analytical chemistry, 94(25).
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