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Dna dye dapi

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DAPI is a fluorescent dye that binds to DNA. It is commonly used in molecular biology and cell biology applications to stain and visualize nucleic acids.

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Lab products found in correlation

Fitc phalloidin, by Merck Group (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Fv1000 confocal laser scanning microscope, by Olympus (1 mentions) Anti mouse alexa fluor 647, by Abcam (1 mentions) Rabbit anti mpo, by Abcam (1 mentions) Tcs sp2 aobs apparatus, by Leica (1 mentions) Prolong gold anti fade media, by Thermo Fisher Scientific (1 mentions) Ventana discovery ultra, by Roche (1 mentions) Discovery inhibitor, by Roche (1 mentions) Tissuefaxs 200 system, by TissueGnostics (1 mentions) Citrate buffer, by Agilent Technologies (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) Dp71 digital camera, by Olympus (1 mentions) Power block, by BioGenex (1 mentions)

6 protocols using dna dye dapi

1

Actin and Nucleus Staining Protocol

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Cells were washed with PBS (pH 7.4), fixed in methanol, rinsed and permeabilized with PBS containing 0.1% Triton X-100. Fixed cells were blocked for 1 h in 3% BSA and incubated with FITC-phalloidin (Sigma) for 30 min and then washed with PBS. The DNA dye DAPI (Molecular Probes) was used as nuclear stain. Images were obtained using a Laser confocal microscopy in 5 high-powered (×1000) fields. Multiple cells were categorized in each experimental point.
Wang S., Wei H, & Zhang S. (2017). Dickkopf-4 is frequently overexpressed in epithelial ovarian carcinoma and promotes tumor invasion. BMC Cancer, 17, 455.
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2

Immunofluorescence Staining Protocol

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Cells were fixed in methanol at room temperature for 20 min and then blocked with normal goat serum for 30 min. The cells were incubated with the primary antibody overnight at 4°C and subsequently with secondary antibody conjugated with Alexa Fluor 488 (Green) dye from Molecular Probes after washed three times in PBT (PBS with 1‰ triton x-100). The DNA dye DAPI (Molecular Probes) was used (blue). Confocal scanning analysis was performed by using a Leika laser confocal scanning microscope in accordance with established methods, utilizing sequential laser excitation to minimize the possibility of fluorescent emission bleed-through.
Li Y., Ke Q., Shao Y., Zhu G., Li Y., Geng N., Jin F, & Li F. (2015). GATA1 induces epithelial-mesenchymal transition in breast cancer cells through PAK5 oncogenic signaling. Oncotarget, 6(6), 4345-4356.
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3

Immunofluorescence Localization of Cellular Proteins

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The cellular localization of proteins was determined by indirect
immunofluorescence. HSG or HSY cells were grown on sterilized glass coverslips
or 8 well chamber slides, fixed in 4% paraformaldehyde, permeabilized in
0.1% Triton X-100, and blocked in 5% normal goat serum-PBS. The
cells were incubated with primary antibodies for 1 hour, washed with PBS three
times, and then incubated with goat anti-mouse or goat anti-rabbit secondary
antibody conjugated with Alexa 488 (green) or Alexa 555 (red) from Molecular
Probes. For actin cytoskeletal staining, phalloidin conjugated with Alexa 488
was used (Molecular Probes). The DNA dye DAPI (Molecular Probes) was used as
nuclear stain (blue). Microscopic analyses were done using an Olympus FV1000
laser scanning confocal microscope in accordance with established methods, using
sequential laser excitation to minimize fluorescence emission bleed-through.
Each image was obtained at the same magnification.
Ohshiro K, & Kumar R. (2015). MTA1 Regulation of ERβ Pathway in Salivary Gland Carcinoma Cells. Biochemical and biophysical research communications, 464(4), 1016-1021.
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4

Immunohistochemical Staining of PsA and OA Synovia

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Three-μm-thick sections in paraffin of human PsA and OA synovia were stained after deparaffination in xilene (5 min, two times), followed by passages in: absolute ethanol (3 min), 95% ethanol in water (3 min), 80% ethanol in water (3 min), 70% ethanol in water, and antigen retrival (5 min at 95°C in 10 mM sodium citrate, pH 6.0). Slides were saturated with blocking buffer (PBS, 0.05% tween 20, 4% BSA) for 1 hour at room temperature. Specimens were stained with a polyclonal rabbit anti-LL37 (Innovagen), rabbit anti-MPO (Abcam), mouse anti-MxA (Novus Bio), monoclonal mouse anti-LL37 (Mab137), polyclonal rabbit anti-human C9 (ATLAS). The following antibodies were used: donkey anti-rabbit IgG AlexaFluor-568 or-647, anti-mouse AlexaFluor-647 and an anti-goat AlexaFluor-488 (Abcam). After washing, slides were mounted in Prolong Gold anti-fade media containing a DNA dye (DAPI) (Molecular Probes). CLSM observations were performed with a Leica TCS SP2 AOBS apparatus, using a 63x/1.40 NA oil objective. Acquisition of images was performed by a Leica confocal software 2.6 (Leica, Germany).
Frasca L., Palazzo R., Chimenti M.S., Alivernini S., Tolusso B., Bui L., Botti E., Giunta A., Bianchi L., Petricca L., Auteri S.E., Spadaro F., Fonti G.L., Falchi M., Evangelista A., Marinari B., Pietraforte I., Spinelli F.R., Colasanti T., Alessandri C., Conti F., Gremese E., Costanzo A., Valesini G., Perricone R, & Lande R. (2018). Anti-LL37 Antibodies Are Present in Psoriatic Arthritis (PsA) Patients: New Biomarkers in PsA. Frontiers in Immunology, 9, 1936.
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5

Quantitative Immunofluorescence Imaging of Tumor Samples

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Patient prostate tissues or mouse tumors were fixed in 10% formalin in PBS and embedded in paraffin. 4μm sections were baked at 60°C for 1 hr and subjected to Immunofluorescence staining using the Ventana Discovery Ultra autostainer with buffers and antibodies from Ventana Medical Systems (Oro Valley, AZ). Briefly slides were deparaffinized using EZ solution, followed by antigen retrieval with Cell Conditioning 1 (CC1) solution (prediluted Tris solution, pH 8.0) for 64 min at 95°C and blocking with Discovery inhibitor (Ventana) for 12 min at RT. The slides were incubated with αCD8 (Ventana) or αKi-67 (Ventana) at 37°C for 28 minutes followed by detection with anti-rabbit-HQ/ αHQ-HRP/ DAB (Ventana) or Red610 kits (Ventana). DNA dye DAPI (Invitrogen) was used to stain the nucleus. Slides was scanned using TissueFAXS 200 system (TissueGnostics, Tarzana, CA). Image was analyzed using QuPath (Edinburgh, UK) with default settings. For all summarized data, 3 random high-power fields were evaluated by automated image analysis and averaged prior to generating a mean for each experimental group.
Wang Y., You S., Su S., Yeon A., Lo E.M., Kim S., Mohler J.L., Freeman M.R, & Kim H.L. (2021). Cholesterol-lowering Intervention Decreases mTOR Complex 2 Signaling and Enhances Antitumor Immunity. Clinical cancer research : an official journal of the American Association for Cancer Research, 28(2), 414-424.
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6

Intra-tumoral Injection of C101 and C154 in GBM-XD456 Xenografts

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GBM-XD456 cells (3 × 105) were injected in the right cerebral hemisphere of athymic nude mice as previously described.30 (link) The tumor bed was injected 10 days later with 1 × 107 PFU of C101, C154 or saline. Mice were euthanized 24 hours post-injection and the tumors harvested, fixed in formalin, and paraffin embedded for IHC and fluorescence microscopy. Tumor sections were deparaffinized, treated in a Citrate buffer (Dako, Carpinteria, CA) to retrieve antigens, incubated in Power Block (BioGenex, Fremont, CA) to block non-specific binding. Sections were then exposed to CA9 (abcam, Cambridge, MA) primary antibody at 1:500 or normal rabbit IgG as a negative control. Cy3 conjugated AffinityPure F(ab)2 fragment donkey anti-rabbit IgG (Jackson ImmunoResearch Laboratories, Suffolk, UK) was used for a secondary antibody. DNA dye DAPI (Invitrogen) was used to counterstain sections. Slides were mounted with Fluoromount-G (Southern Biotech, Birmingham, AL). Photos were taken microscopically with Olympus IX70 connected to a DP71 digital camera (Olympus, Center Valley, PA) and analyzed with software from the manufacturer.
Friedman G.K., Nan L., Haas M.C., Kelly V.M., Moore B.P., Langford C.P., Xu H., Han X., Beierle E.A., Markert J.M., Cassady K.A, & Gillespie G.Y. (2014). γ134.5-Deleted HSV-1 Expressing Human Cytomegalovirus IRS1 Gene Kills Human Glioblastoma Cells as Efficiently as Wild-type HSV-1 in Normoxia or Hypoxia. Gene therapy, 22(4), 348-355.
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