Antifade medium

Manufactured by Beyotime

Sourced in China

Antifade medium is a solution used in fluorescence microscopy to preserve the fluorescence signal of labeled samples during imaging. It helps prevent the fading or bleaching of fluorescent dyes, allowing for longer observation and documentation of the sample.

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Lab products found in correlation

Inverted fluorescent microscope, by Nikon (1 mentions) Donkey anti rabbit igg h l alexa fluor 488, by Abcam (1 mentions) Goat anti mouse igg h l alexa fluor 568, by Abcam (1 mentions) Rabbit anti cd34, by Abcam (1 mentions) 4 6 diamidino 2 phenylindole dapi, by Cayman Chemical (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Cy3 labelled goat anti rabbit igg, by Beyotime (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Imidazole, by Sinopharm (1 mentions) Penicillin streptomycin antibiotic, by Thermo Fisher Scientific (1 mentions) Glycerol, by Sinopharm (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Potassium chloride (kcl), by Sinopharm (1 mentions) Triton x 100, by Beyotime (1 mentions) Sodium chloride (nacl), by Sinopharm (1 mentions) Na2hpo4 12h2o, by Sinopharm (1 mentions) Rabbit polyclonal anti inos, by Santa Cruz Biotechnology (1 mentions) Image pro plus software, by Media Cybernetics (1 mentions)

Close spelling variants of this product

Antifade Mounting Medium with Hoechst 33342 Antifade Mounting Medium containing DAPI Antifade mounting medium Antifade Mounting Medium with DAPI Antifade Polyvinylpyrrolidone Mounting Medium Antifade Polyvinyl Pyrrolidone Medium DAPI-conjugated Antifade Mounting Medium ProLong Antifade mounting medium Antifade Mounting Medium with 4′,6-diamidino-2-phenylindole (DAPI)

5 protocols using antifade medium

Immunofluorescent Characterization of Cells

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Fresh cells were seeded at a suitable density on microscope slides, washed three
times with PBS, fixed with 4% paraformaldehyde for 20 min, and permeabilized by
treating with 0.5% Triton X-100 for another 10 min. The cells were blocked by
treating with 3% bovine serum albumin (BBI, Shanghai, China) for 1 h. Rat
anti-vimentin (1:100; Cell Signaling Technologies, Danvers, MA, USA) and rabbit
anti-CD34 (1:200; Abcam, Cambridge, UK) antibodies were used. The cells were
treated with the antibodies overnight at 4°C. After washing three times with
PBS, the cells were treated with donkey anti-rabbit IgG (H+L) Alexa Fluor 488
(1:1000; Abcam, Cambridge, UK) or goat anti-mouse IgG (H+L) Alexa Fluor 568
(1:1000; Abcam, Cambridge, UK) at 37°C for 1 h, followed by treatment with
4’,6-diamidino-2-phenylindole (DAPI; Cayman Chemical, Ann Arbor, MI, USA). The
slides were fixed in an antifade medium (1:1000; Beyotime, Shanghai, China) and
imaged using an inverted fluorescent microscope (Nikon, Tokyo, Japan).

Huang Y.L., Zhang F.L., Tang X.L, & Yang X.J. (2021). Telocytes Enhances M1 Differentiation and Phagocytosis While Inhibits Mitochondria-Mediated Apoptosis Via Activation of NF-κB in Macrophages. Cell Transplantation, 30, 09636897211002762.

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Immunohistochemical Identification of Telocytes

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Generally, double-labelled immunofluorescence using CD34 in combination with PDGFR alpha or beta, c-kit and vimentin was the available choice for TCs immunodiagnostics 15 (link),29 (link). Representative serial sections were processed for the primary antibodies in pairs (vimentin versus CD34, vimentin versus c-kit): mouse anti-rat monoclonal vimentin (1:100; cat no. BM0135), rabbit anti-rat polyclonal CD34 (1:100; cat no. BA0532), rabbit anti-rat polyclonal c-kit (1:100; cat no. BA0467-1). Then FITC-goat antimouse IgG for vimentin (1:50; cat no. BA1101), CY3-goat anti-rabbit IgG for CD34 (1:50; cat no. BA1032), TRITC-goat anti-rabbit IgG for c-kit (1:50; cat no. BA1090) were added (all from Boster). Finally, counterstained with DAPI (1:50; cat no. C1002) and mounted with antifade medium (1:1000; cat no. p0126; both from Beyotime). Sections were observed under fluorescence microscope (Olympus BX51; Olympus, Tokyo, Japan). Telocytes was the only kind of cell which were double-positive for vimentin and CD34, with typical TCs morphology and intact nuclei 35 (link). The number of TCs was identified in 10 randomly selected microscopic high-power fields per section (40 × 10 original magnification), and statistically analysed in 30 AS-affected and -unaffected sections respectively 29 (link),32 (link).

Yang J., Chi C., Liu Z., Yang G., Shen Z.J, & Yang X.J. (2015). Ultrastructure damage of oviduct telocytes in rat model of acute salpingitis. Journal of Cellular and Molecular Medicine, 19(7), 1720-1728.

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Immunofluorescent Labeling of POU-M2 Protein

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Br-CC-CA was fixed in 4% paraformaldehyde at 25°C for 30 min, washed thrice with 0.3% PBST (PBS containing 0.3% Triton-X 100 (v/v)) and then incubated with anti-POU-M2 (dilution ratio: 1 : 1,000) at 4°C overnight. Next, the samples were incubated with Cy3-labelled goat anti-rabbit IgG (dilution ratio: 1 : 500) (Beyotime) for 2 h followed by washing with PBS thrice and cell nuclei staining with 4′,6-diamidino-2-phenylindole (dilution ratio, 1 : 1000) (Life Technologies, CA, USA). Finally, the samples were fixed in an anti-fade medium (Beyotime) after washing with PBS buffer thrice and then imaged on an Olympus confocal microscopy FV1000 (Tokyo, Japan). The fluorescence signal was excited at the wavelength of 340 nm and 550 nm, respectively.

Cai R., Tao G., Zhao P., Xia Q., He H, & Wang Y. (2022). POU-M2 promotes juvenile hormone biosynthesis by directly activating the transcription of juvenile hormone synthetic enzyme genes in Bombyx mori. Open Biology, 12(4), 220031.

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Neuronal Cell Culture and Assays

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EGCG, thioflavin T (ThT), D₂O, 2′,7′-Dichlorofluorescin diacetate (DCFH-DA), tris-hydroxymethyl aminomethane (Tris), and phenylmethanesulfonyl fluoride (PMSF) were purchased from Sigma and Aldrich, USA. Horse serum (HS), fetal bovine serum (FBS), phosphate buffered saline (PBS), Dulbecco’s modified Eagle’s medium (DMEM), l-glutamine and penicillin/streptomycin antibiotics were purchased from Gibco, USA. Glycerol, imidazole, cupric chloride (CuCl₂∙H₂O), hydrochloric acid (HCl), KH₂PO₄, Na₂HPO₄·12H₂O, KCl, and NaCl were sourced from Sinopharm, China. Annexin V-FITC (fluorescein isothiocyanate, FITC) and propidium podide (PI) were purchased from Dojindo Laboratories. RIPA buffer, 0.01% poly-l-lysine, goat serum, and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Songon Biotech, China. Antifade medium, paraformaldehyde, and Triton X-100 were purchased from Beyotime, China. The PC12 cell was obtained from Shanghai Institutes for Biological Sciences, China.

Teng Y., Zhao J., Ding L., Ding Y, & Zhou P. (2019). Complex of EGCG with Cu(II) Suppresses Amyloid Aggregation and Cu(II)-Induced Cytotoxicity of α-Synuclein. Molecules, 24(16), 2940.

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Quantitative Analysis of iNOS and COX-2

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iNOS and COX-2 were quantitatively measured by single-labelled immunofluorescence. Briefly, sections were incubated with primary anti-rat antibodies: rabbit polyclonal anti-iNOS (1:100; cat no.sc-649); mouse polyclonal anti-COX-2 (1:100; cat no.sc-166475; all from Santa Cruz Biotechnology, Santa Cruz, CA, USA). Then FITC-goat anti-rabbit/mouse IgG (1:100; cat. no. BA1105/BA1101; all from Boster, Wuhan, China) was added. Finally, the sections were mounted with antifade medium (1:1000; cat no. p0126; Beyotime, Shanghai, China). Immunofluorescence intensity was quantitatively measured by laser confocal scanning microscope (TCS-SP2; Leica Lasertechnik, Heidelberg, Germany). Images were converted to greyscale and pixel values of positive-stained areas were used for staining intensity. Total staining intensity per selected area was calculated by multiplying the number of pixels/area with the area mean intensity. At least 500 cells obtained from three separate areas/microscopical fields for iNOS (COX-2, respectively) were analysed for each experimental condition using Image-Pro plus software (version 6.2, Media Cybernetics, Inc, Bethesda, MD, USA) 37 (link).

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