Dapi containing solution

Manufactured by Vector Laboratories

Sourced in United States

DAPI-containing solution is a fluorescent dye that binds to the adenine-thymine (A-T) rich regions of DNA. It is commonly used in fluorescence microscopy and flow cytometry applications to stain and visualize cell nuclei.

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Lab products found in correlation

Alexa fluor 488 conjugated anti mouse and alexa fluor 555 conjugated anti rabbit antibodies, by Thermo Fisher Scientific (1 mentions) Anti occludin antibody, by Abcam (1 mentions) Lsm800 axioobserver, by Zeiss (1 mentions) Axio observer microscope, by Zeiss (1 mentions) Fitc conjugated goat anti mouse igg secondary antibody, by Thermo Fisher Scientific (1 mentions) Mouse anti nf κb p65, by Cell Signaling Technology (1 mentions) Alexa fluor 488 conjugated anti mouse, by Thermo Fisher Scientific (1 mentions) 12 well plates, by Eppendorf (1 mentions) Alexa fluor 555, by Thermo Fisher Scientific (1 mentions) P nf κb ser536, by Cell Signaling Technology (1 mentions) Anti il 1β, by Santa Cruz Biotechnology (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Confocal microscope, by Nikon (1 mentions)

Close spelling variants of this product

DAPI solution (16 citations) DAPI-containing mounting solution (15 citations) 4′,6-diamidino-2-phenylindole (DAPI)-containing mounting solution (8 citations) Vectashield solution containing DAPI (6 citations) Vectashield mounting solution containing DAPI (4 citations) Vectashield solution containing 4′,6-diamidino-2-phenylindole (DAPI; (4 citations) Emulsion oil solution containing 4’,6-diamidino-2-phenylindole (DAPI) (4 citations) Antifade mounting solution containing DAPI (2 citations)

6 protocols using dapi containing solution

Dasatinib Modulates Microglial and Astrocytic Neuroinflammation

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To examine the effect of dasatinib on LPS-induced neuroinflammatory responses, BV2 microglial cells and mouse primary astrocytes were fixed with 4% paraformaldehyde for 10 min, washed three times with 1x PBS, and then incubated with anti-CD11b and anti-COX-2, anti-CD11b and anti-p-AKT, anti-CD11b and anti-p-ERK, or anti-p-STAT3 antibodies in GDB buffer (0.1% gelatin, 0.3% Triton X-100, 16 mM sodium phosphate, pH 7.4, and 450 mM NaCl) overnight at 4 °C. The next day, BV2 microglial cells and mouse primary astrocytes were washed three times with 1x PBS and incubated with Alexa Fluor 488-conjugated anti-mouse and Alexa Fluor 555-conjugated anti-rabbit antibodies (1:200, Molecular Probes, USA) for 1 h at room temperature. The cells were mounted in DAPI-containing solution (Vector Laboratories, CA, USA), and images were captured from a single plane using a confocal microscope (Nikon, Japan) and analyzed using ImageJ software. Samples were analyzed in a blinded manner using 5–10 individual images.

Ryu K.Y., Lee H.J., Woo H., Kang R.J., Han K.M., Park H., Lee S.M., Lee J.Y., Jeong Y.J., Nam H.W., Nam Y, & Hoe H.S. (2019). Dasatinib regulates LPS-induced microglial and astrocytic neuroinflammatory responses by inhibiting AKT/STAT3 signaling. Journal of Neuroinflammation, 16, 190.

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Intestinal Permeability Quantification

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For intestinal permeability status, immunofluorescence using an anti-Occludin antibody (Abcam, Cambridge, UK) was used in paraffin cut sections of jejunum. A Cy3-labeled secondary antibody was used and slides were mounted in a DAPI-containing solution (Vector Laboratories, Burlingame, CA, USA). Three images per section were taken using a confocal immunofluorescent microscope, Zeiss LSM 800 Axio Observer (Zeiss, Thornwood, NY, USA) (magnification 40×), and fluorescence intensity was analyzed with Image J software. Fluorescence measured at the lumen of the colonic tissue was used as background fluorescence to subtract from the epithelial fluorescence values. Therefore, fluorescence density values represented the total protein in the epithelial cells [78 (link)].

Quesada-Vázquez S., Bone C., Saha S., Triguero I., Colom-Pellicer M., Aragonès G., Hildebrand F., del Bas J.M., Caimari A., Beraza N, & Escoté X. (2022). Microbiota Dysbiosis and Gut Barrier Dysfunction Associated with Non-Alcoholic Fatty Liver Disease Are Modulated by a Specific Metabolic Cofactors’ Combination. International Journal of Molecular Sciences, 23(22), 13675.

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Cytoskeletal Dynamics in Fibroblasts

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Fibroblasts grown on 10kPa, 100kPa, 400 kPa, or glass and treated with neomycin or wortmannin were fixed in 10% formalin at 7 hours after barrier removal, probed for focal adhesions with an antibody to phospho-paxillin [Y113] (Abcam, cat. ab32084, Cambridge, MA) in a 1% BSA, 0.1% Tween-20 solution. Cells were then probed with 1:400 rhodamine phalloidin and secondary antibodies (ThermoFisher, Waltham, MA), mounted in a DAPI-containing solution (Vector laboratories, Burlingame, CA) to counterstain for nuclei, and imaged on a Zeiss Axio Observer microscope with AxioVision software. The actin fiber ending closest to the center of the lamella was determined and measurements taken from it and the two to the right and left, making five measurements per cell. The distances from the paxillin at the end of the 5 actin fibers to the lamellar periphery were measured using ImageJ software and averaged for each for the conditions of stiffness and drug treatment (Fig 2B). Cells at the leading edge of the wound closure were selected in an unbiased manner for each condition with about 5 cells in at least 3 cell cultures, yielding approximately 15 cells per condition.

Mkrtschjan M.A., Gaikwad S.B., Kappenman K.J., Solís C., Dommaraju S., Le L., Desai T.A, & Russell B. (2017). Lipid signaling affects primary fibroblast collective migration and anchorage in response to stiffness and microtopography. Journal of cellular physiology, 233(4), 3672-3683.

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NF-κB p65 Translocation Assay in BV2 Cells

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To detect NF-κB p65 translocation, BV2 microglial cells were rinsed twice with PBS and fixed with 4% paraformaldehyde solution for 10 min, permeabilized with 0.1% (v/v) Triton-X100 for 20 min and blocked with 2% (w/v) BSA for 1 h. Cells were then sequentially incubated with mouse anti-NF-κB p65 (Cell Signaling Technology) overnight at 4 °C, and FITC-conjugated goat anti-mouse IgG secondary antibody (Invitrogen) for 45 min. The cells were then mounted in a DAPI-containing solution (Vector Laboratories, CA, USA), and images were captured by fluorescence microscopy (BX-51, Olympus Corp.).

Phan Van T., Huyen Ton Nu Bao T., Leya M., Zhou Z., Jeong H., Lim C.W, & Kim B. (2024). Amlexanox attenuates LPS-induced neuroinflammatory responses in microglial cells via inhibition of NF–κB and STAT3 signaling pathways. Scientific Reports, 14, 2744.

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Immunofluorescent Profiling of BV2 Microglia

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BV2 microglial cells were fixed with 4% paraformaldehyde for 10 min, washed with PBS three times, and then incubated with anti-CD11b and anti-IL-1β or anti-CD11b and anti-COX-2 antibodies in GDB buffer (0.1% gelatin, 0.3% Triton X-100, 16 mM sodium phosphate, pH 7.4, and 450 mM NaCl) overnight at 4 °C. The next day, the cells were washed with PBS three times and incubated with the following secondary antibodies for 1 h at room temperature: Alexa Fluor 488-conjugated anti-mouse and Alexa Fluor 555-conjugated anti-rabbit (1:200, Molecular Probes, USA). The cells were mounted in DAPI-containing solution (Vector Laboratories, CA, USA), and images were captured from a single plane using a confocal microscope (Nikon, Japan) and analyzed using ImageJ software. Samples were analyzed in a blinded manner using 6–10 individual images.

Nam H.Y., Nam J.H., Yoon G., Lee J.Y., Nam Y., Kang H.J., Cho H.J., Kim J, & Hoe H.S. (2018). Ibrutinib suppresses LPS-induced neuroinflammatory responses in BV2 microglial cells and wild-type mice. Journal of Neuroinflammation, 15, 271.

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Visualizing NF-κB Activation in LPS-Treated Cells

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Cells were plated at a density of 20,000 cells/well on cover slips in 12-well plates (Eppendorf, Hamburg, Germany). Twenty four hour after exchanging the culture medium with serum-free high-glucose DMEM (HyClone), the cells were pretreated with ALWPs (250 μg/ml) or PBS for 30 min, followed by treatment with LPS (1 μg/ml) or PBS for 5.5 h. The cells were then fixed in cold methanol for 8 min, washed with PBS three times, and incubated with one of the following primary antibodies in GDB buffer [0.1% gelatin, 0.3% Triton X-100, 16 mM sodium phosphate (pH 7.4), and 450 mM NaCl] overnight at 4°C: anti-CD11b (M1/70, Cat No: ab8878, 1:200, Abcam), anti-IL-1β (Cat No: sc-7884, 1:200, Santa Cruz Biotechnology), or p-NF-κB (Ser536, Cat No: 3033L, 1:100, Cell Signaling Technology). The next day, the cells were washed with PBS three times and incubated with the following secondary antibodies for 1 h at room temperature: Alexa Fluor 488 and Alexa Fluor 555 (1:200, Molecular Probes, OR, United States). The cells were mounted in DAPI-containing solution (Vector Laboratories, CA, United States), and images were acquired on a single plane using a confocal microscope (Nikon, Japan) and analyzed using ImageJ software.

Lee J.Y., Joo B., Nam J.H., Nam H.Y., Lee W., Nam Y., Seo Y., Kang H.J., Cho H.J., Jang Y.P., Kim J., We Y.M., Koo J.W, & Hoe H.S. (2018). An Aqueous Extract of Herbal Medicine ALWPs Enhances Cognitive Performance and Inhibits LPS-Induced Neuroinflammation via FAK/NF-κB Signaling Pathways. Frontiers in Aging Neuroscience, 10, 269.

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