Prolong gold antifade mounting media with dapi

Manufactured by Thermo Fisher Scientific

Sourced in United States

ProLong Gold antifade mounting media with DAPI is a specialized solution used in fluorescence microscopy applications. It is designed to protect fluorescent dyes, such as DAPI, from photobleaching and maintain the signal intensity of fluorescent samples. The product contains an antifade agent and the nuclear stain DAPI, which labels DNA in cell nuclei.

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Lab products found in correlation

Fluorescence conjugated alexa fluor secondary antibodies, by Thermo Fisher Scientific (4 mentions) Lsm 880 confocal microscope, by Zeiss (4 mentions) Anti 3 nitrotyrosine 3 nt, by Abcam (3 mentions) Streptavidin hrp vectastain elite abc kit, by Vector Laboratories (3 mentions) Anti il 1β, by Abcam (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Anti tlr4, by Abcam (2 mentions) Anti occludin, by Abcam (2 mentions) Anti claudin 2, by Abcam (2 mentions) Corticosterone, by Merck Group (2 mentions) Species specific biotinylated conjugated secondary antibodies, by Vector Laboratories (2 mentions) A11008, by Thermo Fisher Scientific (2 mentions) Karyomax colcemid, by Thermo Fisher Scientific (2 mentions) Parpi olaparib, by Selleck Chemicals (2 mentions) Anti α sma, by Abcam (2 mentions) Vectastain elite abc kit, by Vector Laboratories (2 mentions) Lsm 880, by Zeiss (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Xylene, by Thermo Fisher Scientific (2 mentions) Fluorescent labeled secondary antibody, by Thermo Fisher Scientific (2 mentions)

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ProLong Gold Antifade with DAPI (258 citations) ProLong Gold Antifade mounting media (145 citations) Prolong Gold mounting media with DAPI (25 citations) ProLong Gold Antifade DAPI (24 citations) ProLong antifade mounting media (21 citations) ProLong Gold Antifade Mounting with DAPI (7 citations) Prolong Gold DAPI mounting media (6 citations) Prolong Gold Antifade media with DAPI (2 citations) ProLong Mounting Media with DAPI (2 citations) Antifade Mounting media with DAPI (2 citations)

37 protocols using prolong gold antifade mounting media with dapi

Immunofluorescence Staining of DRG and Sciatic Nerve

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Just prior to staining, DRG sections were thawed and dried. All blocking and antibody incubations and washes were performed at room temperature in stationary humid chambers. Sections were incubated in PBT blocking agent (1X PBS, 0.1 % TritonX-100, 1% BSA, 1% donkey serum) for 2 hours, then overnight in primary antibodies diluted in PBT blocking agent. Two sections from each of the four treatment groups (vehicle paclitaxel, paclitaxel, vehicle eribulin and eribulin) were stained simultaneously with the same antibody solution. Stained slides were mounted using ProLong Gold Antifade mounting media with DAPI (ThermoFisher Scientific). Slides were stored at 4°C prior to and after imaging.
Sciatic nerve sections were grouped so that one section from each treatment condition was stained with the same antibody solution. Antibodies were diluted to the same concentrations used in DRG staining in PBT block. Sections were incubated free-floating, at 4°C for 7 days to promote antibody penetration, washed three times in PBT (10 minutes each wash) and then incubated in secondary antibody for two days. Sections were then washed three times, ten minutes each, as in the previous wash step and mounted onto glass slides with ProLong Gold Antifade mounting media with DAPI (ThermoFisher Scientific). Slides were stored at 4°C until imaging.

Benbow S.J., Wozniak K.M., Kulesh B., Savage A., Slusher B.S., Littlefield B.A., Jordan M.A., Wilson L, & Feinstein S.C. (2017). Microtubule Targeting Agents Eribulin and Paclitaxel Differentially Affect Neuronal Cell Bodies in Chemotherapy Induced Peripheral Neuropathy. Neurotoxicity research, 32(1), 151-162.

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Visualizing Subcellular Protein Localization

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Cells were seeded and transfected on glass coverslips. 24 hr post-transfection, cells were fixed with 4% paraformaldehyde in PBS for 15 min, permeabilized by 0.1% Triton X-100 (or 5 μg/ml digitonin as indicated) for 5 min, blocked in PBS containing 2% goat serum and 2% BSA for 30 min, and incubated with primary antibodies in blocking solution for 2 hr at room temperature. After washes, cells were stained with fluorophore-conjugated secondary antibodies (Thermo Fisher) for 1 hr at room temperature, and then further washed. Coverslips were mounted on glass slides in ProLong Gold antifade mounting media with DAPI (Thermo Fisher) and sealed. Pictures were taken by an LSM 710 confocal microscope (ZEISS). Super-resolution images were taken by 3D-SIM using Deltavision OMX (GE Healthcare) or Airyscan using LSM 880 confocal microscope (ZEISS). To determine sub-mitochondrial localization of MOK2, transfected COS-7 cells were stained with 500 nM MitoTracker Red CMXRos and monitored with Hessian SIM microscopy as previously described (Huang et al., 2018 (link)). Time-lapse live-cell imaging was performed with a DeltaVision Elite Cell Imaging System (GE Healthcare).

Zhang H., Cao X., Tang M., Zhong G., Si Y., Li H., Zhu F., Liao Q., Li L., Zhao J., Feng J., Li S., Wang C., Kaulich M., Wang F., Chen L., Li L., Xia Z., Liang T., Lu H., Feng X.H, & Zhao B. (2021). A subcellular map of the human kinome. eLife, 10, e64943.

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Intestinal Barrier Disruption Mechanism

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Neomycin trisulfate salt (Neomycin), Enrofloxacin, Pyridostigmine bromide (PB), Permithrin, lipopolysaccharides (LPS) and Corticosterone were purchased from Sigma- Aldrich (St. Louis, MO). Anti- claudin-2, anti- Occludin, anti-TLR4, anti-flotillin, anti 3-nitrotyrosine (3NT), anti-IL1β, and anti-MCP-1 primary antibodies were purchased from Abcam (Cambridge, MA). Species specific biotinylated conjugated secondary antibodies and Streptavidin-HRP (Vectastain Elite ABC kit) was purchased from Vector Laboratories (Burlingame, CA). Fluorescence conjugated (alexa fluor) secondary antibodies, ProLong Gold antifade mounting media with DAPI and Pierce LAL chromogenic endotoxin quantitation kit was bought from Thermo Fisher Scientific (Waltham, MA). All other chemicals which were used in this project purchased from sigma only if otherwise specified. Paraffin-embedding of tissue sections on slides were done by AML laboratories (Baltimore, MD). Microbiome analysis was done by Second Genome, the microbiome company (San Francisco, CA).

Alhasson F., Das S., Seth R., Dattaroy D., Chandrashekaran V., Ryan C.N., Chan L.S., Testerman T., Burch J., Hofseth L.J., Horner R., Nagarkatti M., Nagarkatti P., Lasley S.M, & Chatterjee S. (2017). Altered gut microbiome in a mouse model of Gulf War Illness causes neuroinflammation and intestinal injury via leaky gut and TLR4 activation. PLoS ONE, 12(3), e0172914.

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Tumor Vascular Characterization in Mice

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Paraffin-embedded tumor sections were stained with hematoxylin and eosin (H&E) for cell morphologic evaluation (Darmstadt, Cat No.PS103-01). Sections were imaged with a BX51 light microscope and analyzed using spot 5.0 software. For analysis of tumor blood vessel were perfused with 2MDa FITC-dextran (green) via a tail-intravenous injection of tumor-bearing mice. For immunofluorescence studies in tumor tissues, fixed frozen sections were blocked with 10% goat serum albumin, and incubated overnight with the following primary antibodies: CD31(1:50, Abcam, ab28364); α-SMA (dilution 1:100; Cat No. M0851 Dako), Ki67 (1:100; Abcam, ab16667), Ve-cadherin (1:50; eBioscience, 14–1449-82) and YY1 (dilution 1:100; Cat No.Ab109231, Abcam). For unconjugated antibodies, appropriate secondary antibodies (dilution 1:500; Sigma-Aldrich, Cat No.ab1507, Taufkirchen) were added on the following day, before mounting with Prolong Gold anti-fade mounting media with DAPI (Thermo Fisher, Cat No.D1306).

Liu H., Qiu Y., Pei X., Chitteti R., Steiner R., Zhang S, & Jin Z.G. (2020). Endothelial specific YY1 deletion restricts tumor angiogenesis and tumor growth. Scientific Reports, 10, 20493.

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MCF-7 and T-47D Cell Response to OTX015 and IR

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Indicated MCF-7 and T-47D cells (1 × 10⁴) were plated on coverslips and after overnight culture, cells were treated with 1 µM OTX015 or DMSO for 24 hours followed by IR (4 Gy). 30 minutes and 120 minutes after IR treatment, cells were fixed in 4% paraformaldehyde, permeabilized using PBS with 0.3% Triton-X, and blocked with 5% goat normal serum. The coverslips were incubated overnight with Ser139 phosphorylated histone H2AX (γH2AX) antibody (1:1000 dilution; Cat #05-636, Millipore), followed by Alexa Fluor 488 conjugated anti-mouse antibody (1:200 dilution; Cat# A-21121, Thermo Fisher Scientific). The nuclei were counterstained and coverslips were mounted using ProLong Gold Antifade Mounting media with DAPI (Thermo Fisher Scientific). Images were captured by fluorescence microscopy (Zeiss, #LSM 880) and foci in the nucleus (at least in 40 nuclei for each treatment group of the experiment) were counted manually and plotted as the number of foci per nucleus.

Udden S.M., Baek G., Pandey K., Vidal C., Liu Y., Rahimi A.S., Kim D.N., Nwachukwu C.R., Mani R.S, & Alluri P.G. (2023). Towards precision radiation oncology: endocrine therapy response as a biomarker for personalization of breast radiotherapy. NPJ Precision Oncology, 7, 11.

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Quantification of Nuclear NFAT1 Expression

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Cells were washed and settled on Shandon Single Cytoslide slides by cytospin at 450 rpm for 5 min. Cells were then fixed with 4% paraformaldehyde (CytoFIx, BD Biosciences) for 20 min at room temperature and then washed in PBS. Cells were then permeabilized with methanol (Perm Buffer III, BD Biosciences), washed in PBS and blocked with Blocking buffer (Sigma) for 30 min at room temperature. Slides were incubated overnight at 4°C with NFAT1 antibody (1:150) (CST). The following day, cells were washed three times with PBS and incubated for 1 hour at room temperature with Anti-rabbit IgG AF-647 secondary antibody (Invitrogen). Slides were washed with PBS, dried and mounted in Prolong Gold anti-fade mounting media with DAPI (ThermoFisher). Images were acquired with an Olympus IX3–55 and analyzed by CellSens imaging software (Olympus). Quantitative image analysis was conducted using QuPath 0.2.2 (Bankhead et al., 2017 (link)). Nuclei were first detected using QuPath’s implementation of StarDist, selecting the DAPI channel for nuclear detection. Objects too small to be cells were automatically removed. Next, manual curation of cells was conducted to remove incorrectly segmented objects. At least 50 cells were scored for each condition. Finally, the median nuclear Cy5 signal was used to determine positivity for NFAT1 within all cells.

Sadras T., Martin M., Kume K., Robinson M.E., Saravanakumar S., Lenz G., Chen Z., Song J.Y., Siddiqi T., Oksa L., Knapp A.M., Cutler J., Cosgun K.N., Klemm L., Ecker V., Winchester J., Ghergus D., Soulas-Sprauel P., Kiefer F., Heisterkamp N., Pandey A., Ngo V., Wang L., Jumaa H., Buchner M., Ruland J., Chan W.C., Meffre E., Martin T, & Müschen M. (2021). Developmental partitioning of SYK and ZAP70 prevents autoimmunity and cancer. Molecular cell, 81(10), 2094-2111.e9.

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Intestinal Barrier Dysfunction Assessment

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Pyridostigmine bromide (PB), Permethrin (Per), Lipopolysaccharides (LPS), Corticosterone and Sodium butyrate (NaBT) were purchased from Sigma-Aldrich (St. Louis, MO). Anti-claudin-2, anti-occludin, anti-TLR4, anti-flotillin, anti-HMGBl, anti-Leptin and anti-IL1β primary antibodies were purchased from Abeam (Cambridge, MA). Anti-TLR5 primary antibody was purchased from Santacruz Biotechnology (Dallas, TX). Species-specific biotinylated conjugated secondary antibodies and Streptavidin-HRP (Vectastain Elite ABC kit) were purchased from Vector Laboratories (Burlingame, CA). Fluorescence-conjugated (Alexa Fluor) secondary antibodies, ProLong Gold antifade mounting media with DAPI were purchased from Thermofisher Scientific (Grand Island, NY) and all other chemicals which were used in this project purchased from sigma only if otherwise specified. Paraffin-embedding of tissue sections on slides were done by AML laboratories (Baltimore, MD). Microbiome analysis was done by Second Genome, the microbiome company (San Francisco, CA).

Seth R.K., Kimono D., Alhasson F., Sarkar S., Albadrani M., Lasley S.K., Horner R., Janulewicz P., Nagarkatti M., Nagarkatti P., Sullivan K, & Chatterjee S. (2018). Increased butyrate priming in the gut stalls microbiome associated-gastrointestinal inflammation and hepatic metabolic reprogramming in a mouse model of Gulf War Illness. Toxicology and applied pharmacology, 350, 64-77.

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Immunofluorescence Staining of Transfected Cells

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Cells were seeded and transfected on glass coverslips, 24 h post-transfection, cells were fixed with 4% paraformaldehyde in PBS for 15 min, permeabilized with 0.1% Triton X-100 for 5 min. cells were blocked and incubated with primary antibodies against HA tag (Rabbit anti-HA C29F4, CST, Cat# 3724, dilution 1:500 v/v) and OLLAS tag (Rat anti-OLLAS L2, Novus Biologicals, Cat# NBP1-06713, dilution 1:100 v/v) for 2 h at room temperature. After washes, cells were stained with indicated fluorophore-conjugated secondary antibodies (Thermo Fisher, A-11008, A-11006) for another 1 h at room temperature. After wash, coverslips were mounted by ProLong Gold antifade mounting media with DAPI (Thermo Fisher, P36941). Images were taken by an LSM 880 confocal microscope (ZEISS).

Li S., Han S., Zhang Q., Zhu Y., Zhang H., Wang J., Zhao Y., Zhao J., Su L., Li L., Zhou D., Ye C., Feng X.H., Liang T, & Zhao B. (2022). FUNDC2 promotes liver tumorigenesis by inhibiting MFN1-mediated mitochondrial fusion. Nature Communications, 13, 3486.

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Metaphase Analysis of 53BP1-Expressing MEFs

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MEF cells derived from Tp53bp1^−/− Brca1^Δ11/Δ11 mice (Bunting et al., 2010 (link)) were infected with 53BP1-expressing retroviruses. At approximately 70% confluence, cells were treated with DMSO or 1 μM of the PARPi olaparib (SelleckChem) for 24 hours. After this treatment, MEFs were arrested in mitosis by incubation with 0.2 μg/mL KaryoMax colcemid (Thermo Scientific) for 1 hour. Cells were then harvested by trypsinization, pelleted by centrifugation, and incubated with 75 mM KCl at 37°C for 30 minutes. After KCl incubation, cells were pelleted and fixed by dropwise addition of 500 μL Carnoy’s fixative (3:1 methanol and acetic acid). The mixture was further resuspended in 10 mL of fixative and stored at 4°C until preparation of slides. For metaphase analysis, fixed cells were pelleted by centrifugation and resuspended in 200-500 μL of fixative. Resuspended cells were dropped on glass coverslips, dried in ambient conditions, and coverslips were mounted using ProLong Gold Antifade mounting media with DAPI (Thermo Scientific).

Setiaputra D., Escribano-Diaz C., Reinert J.K., Sadana P., Zong D., Callen E., Sifri C., Seebacher J., Nussenzweig A., Thomä N.H, & Durocher D. (2022). RIF1 acts in DNA repair through phosphopeptide recognition of 53BP1. Molecular cell, 82(7), 1359-1371.e9.

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Immunofluorescent Staining of Myosin in Cells and Zebrafish

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In cells, the MF20 primary antibody was utilized for detection of myosin heavy chain at a dilution of 1:40 (Developmental Studies Hybridoma Bank; DSHB) in combination with an Alexa Fluor 488 goat anti-mouse IgG (H + L) secondary antibody at a dilution of 1:500 (Thermo). DNA was detected using ProLong Gold Antifade mounting media with DAPI (Thermo). MF20 staining in cells was performed as in Kendall et al., 2012 (link). In zebrafish embryo whole mounts, the MF20 antibody was used for the detection of myosin at a concentration of 1:100 in combination with an anti-GFP polyclonal antibody directly conjugated to Alexa Fluor 488 at 1:500 (Thermo). The secondary antibody used was Alexa Fluor 594 goat anti-mouse IgG (H + L) at 1:500 (Thermo). Images were taken on a Keyence BZ-X700 fluorescent microscope.

Kendall G.C., Watson S., Xu L., LaVigne C.A., Murchison W., Rakheja D., Skapek S.X., Tirode F., Delattre O, & Amatruda J.F. (2018). PAX3-FOXO1 transgenic zebrafish models identify HES3 as a mediator of rhabdomyosarcoma tumorigenesis. eLife, 7, e33800.

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