Slowfade antifade

Manufactured by Thermo Fisher Scientific

Sourced in United States

SlowFade Antifade is a reagent designed to slow the fading of fluorescent signals in microscopy samples. It helps preserve the intensity and longevity of fluorescent labels during imaging.

Automatically generated - may contain errors

Lab products found in correlation

Em grade formaldehyde, by Polysciences (5 mentions) Lsm 5 3.2 image capture and analysis software, by Zeiss (3 mentions) Alexa fluor 647 phalloidin, by Thermo Fisher Scientific (3 mentions) Alexa fluor 555 secondary antibody, by Thermo Fisher Scientific (3 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions) Tcs sp2 confocal laser scanning microscope, by Leica (2 mentions) Np 40, by Merck Group (2 mentions) Sytox green, by Thermo Fisher Scientific (2 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (2 mentions) Rhodamine phalloidin, by Merck Group (2 mentions) Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions) Lsm 510 confocal microscope, by Zeiss (2 mentions) Lsm 510, by Zeiss (2 mentions) Tcs sp2, by Leica (2 mentions) D3571, by Thermo Fisher Scientific (2 mentions) Schneider s media, by Genesee Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Lsm 510 meta, by Zeiss (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by MP Biomedicals (2 mentions) Anti asc, by Santa Cruz Biotechnology (1 mentions)

Close spelling variants of this product

SlowFade Antifade reagent SlowFade Diamond Antifade Mountant with DAPI ToPro3 and SlowFade diamond antifade with DAPI SlowFade antifade solution SlowFade Gold Antifade SlowFade Diamond Antifade Reagent SlowFade Light Antifade kit SlowFade Gold antifade Mountant with DNA-specific counter stain 4′,6-diamidino-2-phenylindole SlowFade Diamond Antifade Mountant SlowFade® Gold antifade medium

27 protocols using slowfade antifade

Dye-Transfer Assay for Connexin-Mediated Communication

Check if the same lab product or an alternative is used in the 5 most similar protocols

HeLa^Cx43 cells were treated with Saracatinib, PF4618433, U0126, or both inhibitors 3 hr prior to PMA (100 nM) treatment. 1 hr after PMA treatment, cell imaging dishes with a 100% confluent monolayer were removed from the incubator. All buffer and medium used for Dye-transfer assay were pre-warmed at 37°C. After two washes with 1x PBS, cells were overlaid with 1x PBS containing Lucifer yellow. Cells were scrape-loaded with a fine edged micro-scalpel by three longitudinal scratches and then incubated at room temperature for 5 min. Cells were rinsed twice with 1x PBS and incubated in cell culture medium at 37°C for 5 min. After incubation, cells were washed twice with 1x PBS containing 1 mM CaCl₂ and 1 mM MgCl₂ to stop dye spread and fixed with 4% Paraformaldehyde for 20 min. After stained with DAPI for 10 min, cells were mounted by adding several drops of SlowFade anti-fade (Invitrogen). Cells were imaged using confocal microscopy.

Zheng L., Trease A.J., Katsurada K., Spagnol G., Li H., Shi W., Duan B., Patel K.P, & Sorgen P.L. (2020). Inhibition of Pyk2 and Src activity improves Cx43 gap junction intercellular communication. Journal of molecular and cellular cardiology, 149, 27-40.

+ Open protocol

+ Expand

Multicolor Immunofluorescence Staining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Deparaffinized and rehydrated sections were immersed in 1 mM EDTA solution, pH 8.0, and microwaved for antigen retrieval. Samples were briefly rinsed in phosphate-buffered saline (PBS) solution, permeabilized with 0.2% Tween in PBS, and blocked with 1% bovine serum albumin and 10% normal serum in PBS. Primary antibodies: mouse anti-CD4 mAb (NCL-L-CD4-1F6, Newcastle upon Tyne, UK), rabbit anti-CD14 (clone EPR3653, Roche), rat anti-CXCL10/IP-10 mAb (clone IR2, provided by Morten Ruhwald), were incubated over night at 4°C. After washing, samples were incubated with Jackson donkey anti-rat Cy3 antibody and Alexa488, Alexa647 fluorochrome-coupled goat secondary antibodies directed against rabbit or mouse. Coverslips were mounted in SlowFade-Anti-Fade (Invitrogen, Life Technologies). Fluorescence was analyzed with a TCS SP2 confocal laser scanning microscope (Leica Microsystems, Wetzlar, Germany). Digital images obtained through a 63× objective (zoom factor 2×) were acquired with Leica Confocal Software.

Blauenfeldt T., Petrone L., del Nonno F., Baiocchini A., Falasca L., Chiacchio T., Bondet V., Vanini V., Palmieri F., Galluccio G., Casrouge A., Eugen-Olsen J., Albert M.L., Goletti D., Duffy D, & Ruhwald M. (2018). Interplay of DDP4 and IP-10 as a Potential Mechanism for Cell Recruitment to Tuberculosis Lesions. Frontiers in Immunology, 9, 1456.

+ Open protocol

+ Expand

Vascular Remodeling Histology and Immunostaining

Check if the same lab product or an alternative is used in the 5 most similar protocols

The arteries were isolated, fixed in 4% paraformaldehyde and then embedded in paraffin. The embedded arteries were cut into 5 mm sections, stained with H&E and then examined by light microscopy. Huawei P20 mobile phone was used to record images.
Formaldehyde-fixed paraffin sections were incubated with primary antibodies including anti-NEK7 (1:100, BioSS), anti-NLRP3 (1:100, CST), anti-ASC (1:100, Santa Cruz) and anti-caspase-1 (1:100, CST) antibodies overnight at 4℃ and then incubated with horseradish peroxidase (HRP)-conjugated secondary antibodies for 1 hour at 37℃. Huawei P20 mobile phone was used to record images.
Additionally, the immunofluorescence technique was applied. After antigen retrieval, the slides were incubated with the primary antibodies overnight at 4°C. Next, an antimouse or antirabbit fluorescein isothicyanate (FITC) or tetramethyl rhodamine isothiocyanate (TRITC)-conjugated secondary antibody (1:100, Invitrogen) was added for 2 hours at 37°C. Then 4',6-diamidino-2-phenylindole (DAPI) was added at a dilution of 1:20 000 for 5 min, and the slides were cover-slipped with SlowFade Antifade (Invitrogen). The images were taken at 100 and 200 power magnification using an ix70 microscope and Magnafire V.1.1 software (both Olympus), and Image Pro-Plus software was used to generate colour composite images.

Cai H., Wang P., Zhang B, & Dong X. (2020). Expression of the NEK7/NLRP3 inflammasome pathway in patients with diabetic lower extremity arterial disease. BMJ Open Diabetes Research & Care, 8(2), e001808.

+ Open protocol

+ Expand

Intracellular Immunostaining of EpCAM, 84-1, and CD45 in Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After CD45 depletion and 84-1 selection, cells were plated on a glass slide with Cytofuge (Iris). For intracellular EpCAM staining, cells were fixed using 4% paraformaldehyde for 15 min, washed with PBS (pH 7.4), and permeabilized in PBS (pH 7.4)/0.2% NP40 (Sigma Aldrich) for 20 min. These cells were then blocked in 10% fetal calf serum (Gibco, Invitrogen) for 1 h, and labeled with EpCAM, 84-1 or CD45 primary antibody (1:100) overnight at 4°C. Cells were then rinsed in PBS (pH 7.4) and stained with Alexa Fluor-555 secondary antibodies (Invitrogen) (1:250) for EpCAM (species: rabbit), Alexa Fluor-647 for 84-1 (species: mouse). For nuclei staining, Sytox Green (Invitrogen) (1:1000) was incorporated along with secondary antibody for 60 min. The cells were then washed with PBS (pH 7.4) three times for 15 min each and mounted in Slow fade antifade (Invitrogen). For confocal analysis, images were acquired in 8 bits with the Zeiss LSM 510 confocal microscope using LSM 5 3.2 image capture and analysis software (Zeiss). A 63× water-immersion objective lens (NA, 1.0) was used with digital zoom for image capture. All images were acquired by the same operator using the same intensity and photo detector gain in order to allow quantitative comparisons of relative levels of immunoreactivity between different samples.

Satelli A., Brownlee Z., Mitra A., Meng Q.H, & Li S. (2014). Circulating tumor cell enumeration using a combination of EpCAM and Cell-surface vimentin based methods for monitoring breast cancer therapeutic response. Clinical chemistry, 61(1), 259-266.

+ Open protocol

+ Expand

Immunofluorescence Analysis of Organelle Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

The 2fTGH cells were fixed in ice-cold ethanol/acetone (1:1) at −20 °C for 10 min. Samples were briefly rinsed in phosphate-buffered saline (PBS), permeabilized with 0.1 % Triton X-100 in PBS for 5 min and blocked with 1 % bovine serum albumin (BSA) and 10 % normal goat serum in PBS for 30 min. Primary antibodies, namely mouse anti-TG2 (CUB 7402, Thermo Scientific, Rockford, Ill., USA), rabbit anti-calreticulin antibody (Stressgen, Enzo Life Sciences, Farmingdale, N.Y., USA), rabbit anti-p62/SQSTM1 (MBL, Woburn, Mass., USA) and rabbit anti-Tom20 antibody (Santa Cruz Biotechnology, Dallas, Texas, USA), were incubated for 1 h with the cells at room temperature. After being washed, the cells were incubated with Alexa488- or Alexa594-fluorochrome-coupled secondary antibodies directed against rabbit or mouse (Molecular Probes, Life Technologies, Grand Island, N.Y., USA).
Coverslips were mounted in SlowFade-Anti-Fade (Invitrogen, Life Technologies). Fluorescence was analyzed with a TCS SP2 confocal laser scanning microscope (Leica Microsystems, Wetzlar, Germany). Digital images obtained separately in both channels through a 63× objective (zoom factor 2×) were acquired with Leica Confocal Software.

Piacentini M., D’Eletto M., Farrace M.G., Rodolfo C., Del Nonno F., Ippolito G, & Falasca L. (2014). Characterization of distinct sub-cellular location of transglutaminase type II: changes in intracellular distribution in physiological and pathological states. Cell and Tissue Research, 358(3), 793-805.

+ Open protocol

+ Expand

Immunostaining Drosophila Ovarian Egg Chambers

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Ovaries were dissected in S2 media warmed to room temperature and fixed in PBS +0.1% Triton + 4% EM-grade formaldehyde (Polysciences). For actin staining only, ovaries were washed 3x in PBS + 0.1% Triton (PBT) then egg chambers were disrupted from the muscle sheath by gentle pipetting. Staining was performed with rhodamine phalloidin (1:200, Sigma), AlexaFluor-488 Phalloidin (1:200, Invitrogen) or AlexaFluor-647 Phalloidin (1:75, Invitrogen) for 15 minutes in PBT at room temperature while rocking. Egg chambers were then washed 3x in PBT and mounted in SlowFade Antifade (Invitrogen). Egg chambers stained with primary antibodies were incubated overnight at 4°C with rocking in PBT + 0.1% BSA + Phalloidin. Egg chambers were incubated in AlexaFluor-488 or 555 conjugated secondary antibodies (1:200, Invitrogen) in PBT + 0.1% BSA + Phalloidin for 3 hours at room temperature with rocking. The following primary antibodies were used: anti-Ena (1:200 concentrate, DHSB), anti-SCAR (1:300)^{43 (link)}, and anti-SCAR (1:200 concentrate, DSHB)^{44 (link)}. For SCAR staining, primary and secondary incubations along with all washes following the primary incubation, PBT3 (PBS + 0.3% Triton) was used instead of PBT. Fixed egg chambers were imaged with a laser-scanning confocal or a spinning disk confocal (described above). ImageJ and Adobe Photoshop were used for image processing.

Cetera M., Ramirez-San Juan G.R., Oakes P.W., Lewellyn L., Fairchild M.J., Tanentzapf G., Gardel M.L, & Horne-Badovinac S. (2014). Epithelial rotation promotes the global alignment of contractile actin bundles during Drosophila egg chamber elongation. Nature communications, 5, 5511.

+ Open protocol

+ Expand

Multiprobe FISH Assay for Telomeres and Centromeres

Check if the same lab product or an alternative is used in the 5 most similar protocols

TelG-Cy5 PNA probe in hybridization buffer I (50% deionized formamide, 10% dextran sulfate, 1X SSC, 0.1 μg/ml herring sperm DNA) was pipetted onto prepared slides and spread over the thin layer of agarose gel with a microscope slide coverslip (24 × 60 mm). Slides were placed in a humidified chamber and hybridized at 37°C overnight. The next day, the slides were cooled in cold Wash Solution I (50% formamide, 2X SSC, pH 7.0) and coverslips were gently peeled off. Slides were washed three times with cold Wash Solution I, followed by three more washes with cold 0.1X SSC. Slides were serially dehydrated in 70, 90 and 100% ethanol and allowed to fully dry. These steps were repeated with the TelC-Rho and CENPB-FAM PNA probes together in hybridization buffer II (50% deionized formamide, 10% dextran sulfate, 1X SSC). Slides were counterstained with 0.1 μg/ml 4′, 6-diamidino-2-phenylindole (DAPI) for 5 min and mounted in SlowFade Antifade (Invitrogen) with microscope slide coverslips (24 × 60 mm).

Komosa M., Root H, & Meyn M.S. (2015). Visualization and quantitative analysis of extrachromosomal telomere-repeat DNA in individual human cells by Halo-FISH. Nucleic Acids Research, 43(4), 2152-2163.

+ Open protocol

+ Expand

Immunofluorescence Microscopy Protocol for Protein Labeling

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence microscopy was performed essentially, as described^{67 (link)}. Cells were grown to exponential phase and fixed with 1.6–2.6 % (w/v) paraformaldehyde and 0.008 % (w/v) glutaraldehyde. After permeabilization in GTE buffer (20 mM Tris/HCl, pH 7.6, 50 mM glucose, 10 mM EDTA), the fixed cells were incubated with suitable antibodies in PBS buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na₂HPO₄, 2 mM KH₂PO₄) containing 2 % (w/v) bovine serum albumin (BSA; Carl-Roth, Germany). First, target proteins were labeled with a polyclonal anti-BacO or anti-BacP^{49 (link)} antibody or a monoclonal anti-HA antibody (Millipore) at dilutions of 1:500, 1:400, and 1:200, respectively. Immunocomplexes were then visualized with Alexa-Fluor 594 Goat Anti-Rabbit or Alexa-Fluor 488 Goat Anti-Rabbit secondary antibodies (Molecular Probes) at a dilution of 1:200. Before imaging, SlowFade® Antifade (Invitrogen) was applied to each sample.

Lin L., Osorio Valeriano M., Harms A., Søgaard-Andersen L, & Thanbichler M. (2017). Bactofilin-mediated organization of the ParABS chromosome segregation system in Myxococcus xanthus. Nature Communications, 8, 1817.

+ Open protocol

+ Expand

Immunofluorescence Imaging of Mitochondria

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were cultured on Lab-Tek chambered Slides (#154453, Thermo Fisher Scientific, Hudson, NH) washed with phosphate buffered saline (PBS) and fixed with 4% paraformaldehyde followed by permeabilization with 0.25% Triton X-100. Blocking was done with PBS containing 10% FBS and 0.1% TX-100. Primary antibody incubations were performed in blocking solution at 1:100 dilution over night at 4°C followed by wash with 0.1% TX-100in PBS. Cells were incubated with Alexafluor-conjugated secondary antibodies (Molecular probes, Grand Island, NY) at 1:400 dilutions at room temperature for 1 h. Washed cells were mounted on slides with Slowfade anti-fade (Invitrogen, Grand Island, NY, #S36938) reagent containing DAPI as nuclear stain. For live cell microscopy cells were cultured on Lab-Tek chambered coverglass (Thermo Fisher Scientific, #155383) and stained with 5 µM MitoSox red from Molecular probes (# M36008). Finally cells were observed on a laser scanning confocal microscope (Carl Zeiss).

Mahato B., Home P., Rajendran G., Paul A., Saha B., Ganguly A., Ray S., Roy N., Swerdlow R.H, & Paul S. (2014). Regulation of Mitochondrial Function and Cellular Energy Metabolism by Protein Kinase C-λ/ι: A Novel Mode of Balancing Pluripotency. Stem cells (Dayton, Ohio), 32(11), 2880-2892.

+ Open protocol

+ Expand

Immunofluorescence Staining of Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

For immunofluorescence, the cell pellet was mixed with MACS buffer (Miltenyi Biotec) and stained with 84-1 antibody (1:200) in tube at room temperature for 1 hour. Cells were cytospun onto Polysine^™ microscope adhension slides (Thermo Fisher Scientific) by Cytofuge (Iris, Westwood, MA). Cells were fixed with 4% paraformaldehyde (Fisher Scientific) and blocked in blocking buffer (1% FBS in PBS) for 1 hour. For the staining of other markers, such as CD45 (1:100; Abcam, Cambridge, MA), α-SMA (1:100; Abcam), CD117 (1:100; Cell Signaling Technology, Danvers, MA), and MDM2 (1:100; Santa Cruz Biotechnology Santa Cruz, CA), the cells were incubated with primary antibody overnight in a cold room followed by permeabilization in PBS (pH 7.4)/0.2% NP40 (Sigma Aldrich) for 20 minutes. Next, the slides were washed with PBS three times and stained with Alexa Fluor-555 secondary antibody (1:100; Invitrogen, Carlsbad, CA) for CD45, Alexa Fluor-647 (1:100; Invitrogen) for 84-1, and Sytox Green (1:200; Invitrogen) for nuclei staining for 1 hour at room temperature. Then the slides were washed with PBS three times and mounted in Slow fade antifade (Invitrogen). All slides were visualized by Zeiss LSM 510 confocal microscope using LSM 5 3.2 image capture and analysis software (Carl Zeiss, Thornwood, NY).

Li H., Meng Q.H., Noh H., Batth I.S., Somaiah N., Torres K.E., Xia X., Wang R, & Li S. (2017). Detection of circulating tumor cells from cryopreserved human sarcoma peripheral blood mononuclear cells. Cancer letters, 403, 216-223.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!