Prolong diamond antifade containing dapi

Manufactured by Thermo Fisher Scientific

ProLong Diamond Antifade containing DAPI is a mounting medium designed for fluorescence microscopy. It is formulated to protect fluorescent signals from photobleaching and provides a clear, permanent seal for microscope slides.

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Lab products found in correlation

Alexa fluor 633 goat anti rabbit igg, by Thermo Fisher Scientific (2 mentions) Superfrost plus microscope slide, by Thermo Fisher Scientific (2 mentions) Neg 50, by Thermo Fisher Scientific (2 mentions) Dfc350fx, by Leica (1 mentions) Axio imager a2, by Zeiss (1 mentions) Dm6b widefield fluorescent microscope, by Leica (1 mentions) Metamorph, by Molecular Devices (1 mentions) Csu x1 spinning disk head, by Yokogawa (1 mentions) Alexa 546, by Thermo Fisher Scientific (1 mentions) Phalloidin 488, by Thermo Fisher Scientific (1 mentions) Microm cryostat, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

ProLong Diamond antifade mountant containing DAPI ProLong Diamond Antifade Mountant containing 4′6-diamidino-2-phenylindole (DAPI) Prolong Diamond antifade reagent containing DAPI ProLong™ Diamond containing DAPI ProLong Antifade containing DAPI ProLong Diamond Antifade ProLong DAPI Diamond Prolong Diamond Prolong Antifade DAPI ProLong

5 protocols using prolong diamond antifade containing dapi

Immunofluorescence Evaluation of Autophagy

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Cells were seeded in coverslips dishes at a density of 1 × 10⁴ cells/cm², maintained 24 h for attachment and then treated with BSHE extract (whose GR₅₀ concentrations were those referred to the 48 h-exposure).
After 48 h-incubation, cells were washed twice with PBS, fixed with 4% paraformaldehyde and permeabilized with PBS containing 0.5% Triton X-100. Non-specific binding sites were blocked following a 30 min-exposure to 3% bovine serum albumin (BSA) in PBS-T (PBS and 0.05% Tween-20). Afterwards, cells were incubated with rabbit anti-LC3B antibody (dilution 1:200 in PBS and 1% BSA) at 4°C overnight. Cells were then washed three time with PBS-T and incubated with secondary Dy Light 488-conjugated anti-rabbit antibodies (dilution 1:1,000 in PBS and 1% BSA) in the dark at 4°C for 1 h and washed again with PBS-T.
Coverslips (overturned on microscope slides) were mounted with Pro Long Diamond Antifade containing DAPI (4′,6-Diamidino-2-Phenylindole, Dihydrochloride) (Thermo Fisher Scientific) and observed under fluorescence microscopy (Axio Imager A2 Zeiss). Images were acquired using a black and white camera (Leica, DFC350FX).

Volpe A.R., Carmignani M, & Cesare P. (2023). Hydroalcoholic extract of Buxus sempervirens shows antiproliferative effect on melanoma, colorectal carcinoma and prostate cancer cells by affecting the autophagic flow. Frontiers in Pharmacology, 14, 1073338.

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Aβ Oligomers Impact Na/K ATPase Localization

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Cells matured for 16 DIV on glass coverslips coated with poly-D-lysine
(Sigma P6407) were exposed to AβOs (200, 500 nM) or vehicle for 1
h/37°C. Cells were fixed and labeled as above with 0.1% (v/v) Triton
X-100 in the block for membrane permeabilization (4 h/RT). Primary antibody was
anti-Na/K ATPase-α3 (H-4, mouse mAb, RRID:AB_10848453; 4 μg/mL in
permeabilization buffer) with an Alexa-linked secondary (Sigma A11029).
Coverslips were mounted to glass slides with ProLong Diamond Antifade containing
DAPI (ThermoFisher P36962), air-dried, and imaged using a Leica Spinning Disk
Autofocus Confocal System with a CSU-X1 spinning disk head (Yokogawa Electric
Corporation), a 63x Plan-Apo objective with 1.4 NA (Leica), and an Evolve 512
Delta EMCCD camera (Photometrics). Images were captured with Metamorph
(Molecular Devices; RRID:SCR_002368) and deconvolved using AutoQuant X3’s
(Media Cybernetics; RRID:SCR_002368) iterative, constrained 3D deconvolution
method. Alternatively, images were captured on a Leica DM6B Widefield
Fluorescent Microscope and 3D deconvoluted using Leica software. Images were
thresholded to remove background; remaining pixel area quantified and particles
analyzed in Fiji. Measurements were normalized to cell count. All imaging and
analysis was performed blinded to treatments.

Cline E.N., Das A., Bicca M.A., Mohammad S.N., Schachner L.F., Kamel J.M., DiNunno N., Weng A., Paschall J.D., Bu R.L., Khan F.M., Rollins M.G., Ives A.N., Shekhawat G., Nunes-Tavares N., de Mello F.G., Compton P.D., Kelleher N.L, & Klein W.L. (2019). A Novel Crosslinking Protocol Stabilizes Amyloid β Oligomers Capable of Inducing Alzheimer’s-Associated Pathologies. Journal of neurochemistry, 148(6), 822-836.

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Visualization of Neutrophil Elastase in NET-macrophage Interaction

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Neutrophils (5 × 10⁵/well in Permanox chamber slides (Nalge Nunc) were stimulated with IL-8 and immediately fixed with 4% formaldehyde (FA, Merck) for 30 min and rinsed three times with PBS. Next, samples were blocked with human serum for 1 h and incubated with anti-elastase monoclonal antibodies (1:250; Calbiochem) for two hours. Samples were washed and incubated with secondary antibodies coupled to Alexa546 (Life Technologies) for 30 min, rinsed twice with PBS and mounted with ProLong Diamond anti-fade containing DAPI (Life Technologies). For NET-macrophage interaction assays, these cells were exposed to NETs (500 ng/mL) for 30 min, fixed with FA (4%, 30 min), washed and blocked as described above. Samples were then stained with anti-elastase and Alexa 546-labeled secondary antibodies and Phalloidin-488 (Invitrogen) for 30 min, and then mounted on medium containing DAPI, as above. All staining processes were performed at room temperature (RT). Slides were examined using confocal (Leica DMi8) and fluorescence (Zeiss Ax10) microscopes, and LasX and AxioVision softwares, respectively. Brightness, contrast and color of digital images were adjusted with Adobe Photoshop CS5 v12.0 program (Adobe Systems Inc.).

Mojoli A., Gonçalves B.S., Temerozo J.R., Cister-Alves B., Geddes V., Herlinger A., Aguiar R.S., Pilotto J.H., Saraiva E.M, & Bou-Habib D.C. (2020). Neutrophil extracellular traps from healthy donors and HIV-1-infected individuals restrict HIV-1 production in macrophages. Scientific Reports, 10, 19603.

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Immunohistochemistry for Tyrosine Hydroxylase

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As previously described (Riessland et al., 2019 ), three weeks post injection mice were anesthetized with pentobarbital and transcardially perfused using PBS (pH 7.4), followed by 4% paraformaldehyde in PBS. Brains were postfixed in 4% paraformaldehyde in PBS at room temperature for 1 hour and then cryopreserved using a gradient of 5%, 15% and 30% sucrose. The brains were embedded in Neg-50 (Thermo Scientific), frozen and stored at −80 °C, and cut into 14-μm-thick coronal sections using a Microm cryostat and thaw-mounted onto Superfrost Plus microscope slides (Thermo Scientific). For staining of TH, brain sections were washed in PBS and permeabilized with 0.2% Triton X-100 in PBS, followed by blocking with 2% donkey serum and 0.1% fish gelatin in 0.2% Triton X-100 in PBS. Sections were then incubated with rabbit polyclonal anti-TH antibody (Millipore, #AB152) at a concentration of 1:250 overnight. The next day, slides were washed with PBS, incubated with Alexa Fluor 633 goat anti–rabbit IgG (Life Technologies, #A21070) for 2 hours at room temperature, and then after another washing step were mounted with ProLong Diamond Antifade containing DAPI (Life Technologies, #P36962), coverslipped, and stored in the dark until imaging.

Russo T., Kolisnyk B., Aswathy B., Wan Kim T., Martin J., Plessis-Belair J., Ni J., Pearson J.A., Park E.J., Sher R.B., Studer L, & Riessland M. (2023). The SATB1-MIR22-GBA axis mediates glucocerebroside accumulation inducing a cellular senescence-like phenotype in dopaminergic neurons. bioRxiv.

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Immunohistochemical Analysis of Tyrosine Hydroxylase

Cited 1 time

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As previously described (Riessland et al., 2019 (link)), 3 weeks post injection mice were anesthetized with pentobarbital and transcardially perfused using PBS (pH 7.4), followed by 4% paraformaldehyde in PBS. Brains were postfixed in 4% paraformaldehyde in PBS at room temperature for 1 h and then cryopreserved using a gradient of 5%, 15%, and 30% sucrose. The brains were embedded in Neg‐50 (Thermo Scientific), frozen and stored at −80°C, and cut into 14‐μm‐thick coronal sections using a Microm cryostat and thaw‐mounted onto Superfrost Plus microscope slides (Thermo Scientific). For staining of TH, brain sections were washed in PBS and permeabilized with 0.2% Triton X‐100 in PBS, followed by blocking with 2% donkey serum and 0.1% fish gelatin in 0.2% Triton X‐100 in PBS. Sections were then incubated with rabbit polyclonal anti‐TH antibody (Millipore, #AB152) at a concentration of 1:250 overnight. The next day, slides were washed with PBS, incubated with Alexa Fluor 633 goat anti–rabbit IgG (Life Technologies, #A21070) for 2 h at room temperature, and then after another washing step were mounted with ProLong Diamond Antifade containing DAPI (Life Technologies, #P36962), coverslipped, and stored in the dark until imaging.

Russo T., Kolisnyk B., B. S. A., Plessis‐Belair J., Kim T.W., Martin J., Ni J., Pearson J.A., Park E.J., Sher R.B., Studer L, & Riessland M. (2024). The SATB1‐MIR22‐GBA axis mediates glucocerebroside accumulation inducing a cellular senescence‐like phenotype in dopaminergic neurons. Aging Cell, 23(4), e14077.

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