Triton x 100

Manufactured by Vector Laboratories

Sourced in United Kingdom, Canada, United States

Triton X-100 is a nonionic detergent commonly used in biological research applications. It functions as a surfactant, facilitating the solubilization and extraction of proteins and other biomolecules from cells and tissues.

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Lab products found in correlation

Triton x 100, by Merck Group (3 mentions) Vectashield, by Vector Laboratories (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Triton x 100, by GE Healthcare (1 mentions) Fitc conjugated isolectin b4, by Vector Laboratories (1 mentions) Axio imager m1 microscope, by Zeiss (1 mentions) Sodium metaperiodate, by Merck Group (1 mentions) Triton x, by Thermo Fisher Scientific (1 mentions) Biotinylated goat anti mouse, by Vector Laboratories (1 mentions) Vectastain abc kit, by Vector Laboratories (1 mentions) Alexa fluor 633 phalloidin, by Thermo Fisher Scientific (1 mentions) Bovine serum albumin (bsa), by Vector Laboratories (1 mentions) M o m kit, by Vector Laboratories (1 mentions) Normal horse or goat serum, by Vector Laboratories (1 mentions) Cy5 conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions) Anti neun, by Synaptic Systems (1 mentions) Anti gfap, by Agilent Technologies (1 mentions) Dmi8 platform, by Leica camera (1 mentions) Biotinylated ib4, by Vector Laboratories (1 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions)

8 protocols using triton x 100

Visualizing Laser-Induced Choroidal Neovascularization

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After euthanasia on day 1, 3, 7, or 14 following laser photocoagulation, the eyes were enucleated and prefixed with 4% paraformaldehyde (Nacalai tesque, Kyoto, Japan) for 30 min. Retina-RPE-choroid complexes were microsurgically isolated from the prefixed eyes and further fixed with 4% paraformaldehyde for 1 h. The retina was removed from RPE-choroid complexes. The RPE-choroid complexes were then washed with PBS (1X; 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na₂HPO₄, 1.5 mM KH₂PO₄:Nacalai tesque) and incubated for 30 min with PBS blocking buffer containing 0.5% Triton X-100 (GE Healthcare UK, Buckinghamshire, UK) and 1% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO). To visualize neovascularization, RPE-choroid complexes were incubated overnight at 4 °C with 0.01% fluorescein isothiocyanate (FITC)-conjugated isolectin B4 derived from Griffonia (Bandeiraea) simplicifolia agglutinin (Vector Laboratories, Peterborough, UK) diluted with PBS containing 0.5% Triton X-100. After the RPE-choroid complexes were washed with PBS and sealed with VECTASHIELD (Vector Laboratories), CNV was observed with a fluorescence microscope (IX71; Olympus, Tokyo, Japan). Note that FITC-labeled isolectin binds to microglial cells and macrophages as well as endothelial vascular cells [19 (link)].

Nakajima T., Hirata M., Shearer T.R, & Azuma M. (2014). Mechanism for laser-induced neovascularization in rat choroid: Accumulation of integrin α chain-positive cells and their ligands. Molecular Vision, 20, 864-871.

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Immunofluorescence Staining of BCa Cells

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BCa cells grown on coverslips were fixed with 4% formaldehyde diluted in phosphate buffered saline (PBS) at room temperature (RT) for 15 minutes. After a thorough rinse, cells were incubated with blocking buffer containing 1× PBS, 5% normal donkey serum and 0.3% Triton X-100 (Vector Lab, Burlingame, CA) at RT for 60 minutes, followed by incubation with the primary antibodies (S3 Table) at 4°C overnight. On the second day, cells were incubated with fluorochrome-conjugated second antibody for 1-2 hours at RT in the dark. Final immunofluorescence was observed under an inverted microscope (Axio Imager M1 microscope, Zeiss, Gottingen, Germany).

Xu Y.Y., Yu H.R., Sun J.Y., Zhao Z., Li S., Zhang X.F., Liao Z.X., Cui M.K., Li J., Li C, & Zhang Q. (2018). Upregulation of PITX2 Promotes Letrozole Resistance Via Transcriptional Activation of IFITM1 Signaling in Breast Cancer Cells. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 51(2), 576-592.

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Immunohistochemical Analysis of Hippocampal Tissue

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Twenty-μm-thick sections were cut from paraffin embedded hippocampus (NCI = 9, MCI = 10 and AD = 6) and immunostained (Table 3). After antigen-retrieval in 0.01 M citric acid (pH 8.5) for 15 minutes, sections were washed in phosphate buffer and TBS before a 20-minute incubation in 0.1 M sodium metaperiodate (Sigma) in TBS to inactivate endogenous peroxidase activity. Tissue was permeabilized in TBS containing 0.25% Triton-X (ThermoFisher, Waltham, MA) and blocked in the same solution containing 3% goat serum for 1 hour. Sections were incubated with appropriate antibody dilutions (Table 3) overnight at room temperature in 0.25% Triton X-100, 1% goat serum solution in a wet-chamber, then washed in TBS containing 1% goat serum prior to incubation with the secondary antibody biotinylated goat anti-mouse at a 1:200 dilution for 1 hour (Vector Laboratories, Burlingame, CA). Following TBS washes, sections were incubated using the Vectastain ABC kit (Vector Laboratories) for 1 hour, rinsed in 0.2 M sodium acetate, 1.0 M imidazole buffer, pH 7.4, and developed in acetate-imidazole buffer containing 0.05% 3,3′-diaminobenzidine tetrahydrochochloride (Sigma). Reaction was terminated in acetate-imidazole buffer and slides were dehydrated through graded alcohols (70%-95%-100%), cleared in xylene and cover slipped using DPX (Biochemica Fluka, Buchs, Switzerland).

Perez S.E., He B., Nadeem M., Wuu J., Ginsberg S.D., Ikonomovic M.D, & Mufson E.J. (2015). Hippocampal Endosomal, Lysosomal and Autophagic Dysregulation in Mild Cognitive Impairment: Correlation with Aβ and Tau Pathology. Journal of neuropathology and experimental neurology, 74(4), 345-358.

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Immunohistochemistry and Immunofluorescence for COUP-TFII_V2

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IHC and IF with antibodies to COUP-TFII_V1 (cat. no. AB41859; RRID:AB_742211) and cytokeratin (CK)19 (cat. no. AB15463; RRID:AB_2281021; both from Abcam) were performed as previously described (18 (link),24 (link)). The custom-made primary antibody for COUP-TFII_V2 was used at 1:25 dilution in PBS containing 2% BSA (MilliporeSigma) and 0.01% Triton X-100 (Sigma). IHC for α-smooth muscle actin was performed with the monoclonal clone A4 (DAKO; Agilent Technologies, Inc.) diluted 1:100 in PBS with 2% BSA and 0.01% Triton X-100 following the previously described protocol; the sections were incubated with an M.O.M. kit (Vector Laboratories, Inc.) prior to incubation with the primary antibody. For β-catenin IF the same antibody used in western blot was used diluted 1:50; SMAD2/3P IF was performed with a 1:100 dilution of a rabbit polyclonal antibody (cat. no. ab272332; Abcam); actin filaments were decorated with Alexa Fluor 633 phalloidin (Molecular Probes cat. no. A22284); cell nuclei were stained with DAPI (cat. no. 10236276001; Roche). Collagen was stained with standard Sirius Red staining. Sirius Red was quantified with ImageJ 1.52r under Icy 2.0.3 on images with RGB color settings; the selection of stained areas was achieved with the function 'Color threshold'.

Polvani S., Pepe S., Tempesti S., Tarocchi M., Marroncini G., Bencini L., Ceni E., Mello T., Picariello L., Simeone I., Grappone C., Dragoni G., Antonuzzo L., Giommoni E., Milani S, & Galli A. (2022). Isoforms of the orphan nuclear receptor COUP-TFII differentially modulate pancreatic cancer progression. International Journal of Oncology, 60(5), 55.

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Hippocampal Immunohistochemistry Protocol

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To determine optic fiber and electrode positions, to visualize the hippocampal anatomy after AAV9 injections, and to validate hippocampal sclerosis in ihpKA mice, we performed immunohistochemistry with markers for neurons and astrocytes. For the immunofluorescence staining, free-floating sections were pre-treated with 0.25% TritonX-100 and 10% normal horse or goat serum (Vectorlabs, Burlingame, CA, USA) diluted in PB for 1 h. Subsequently, slices were incubated either with guinea-pig anti-NeuN (1:500; Synaptic Systems, Göttingen, Germany, 266004, RRID:AB_2713971) or rabbit anti-GFAP (1:500, Dako, Glostrup, Denmark, Z0334, RRID:AB_10013382) overnight at 4 °C. Sections were rinsed and then incubated for 2.5 h in donkey anti-guinea-pig or donkey anti-rabbit Cy5-conjugated secondary antibody (1:200, Jackson ImmunoResearch Laboratories Inc., West Grove, PA, USA, anti-guinea-pig: 706-175-148, RRID:AB_2340462; anti-rabbit: 711-175-152, RRID:AB_2340607), followed by extensive rinsing in PB. The sections were mounted on glass slides and coverslipped with Immu-Mount^TM mounting medium (Thermo Shandon Ltd, Runcorn, UK).

Kleis P., Paschen E., Häussler U., Bernal Sierra Y.A, & Haas C.A. (2022). Long-term in vivo application of a potassium channel-based optogenetic silencer in the healthy and epileptic mouse hippocampus. BMC Biology, 20, 18.

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Intragastric Recombinant N1 Decoy Injection in Mice

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C57BL/6 mice postnatal day 1 (P1) pups were injected intragastrically with 12.5 mg/kg of recombinant N1_10-14Fc decoy or Fc for three days (P1-P3). Eyes were isolated at P5 and were fixed in 4% paraformaldehyde (Thermo Fisher Scientific) for 1 h at 4 °C on a nutator. Following fixation, eyes were washed with 1 × PBS solution. Retinas were dissected and permeabilized in 1 × PBS containing 1% BSA (Fisher Bioreagents) and 0.5% Triton X-100 (Fisher Bioreagents) overnight at 4 °C on a nutator. Samples were then immunostained in PBLEC (5% Triton X-100, 1 M MgCl₂, 1 M CaCl₂, and 1 M MnCl₂ in 1 × PBS) overnight at 4 °C with Biotinylated IB4 (1:50; Vector Laboratories, B-1205) and anti-α-SMA-FITC (1:200; MilliporeSigma, F3777). IB4 was detected with streptavidin-conjugated Alexa Fluor 647 (Invitrogen). Immunostained retinas were postfixed with 4% formaldehyde and mounted in Vectashield (Vector Laboratories). Whole-mount retina images were acquired using Leica Dmi8 Platform. All images were analyzed using ImageJ (NIH).

Sargis T., Youn S.W., Thakkar K., Naiche L.A., Paik N.Y., Pajcini K.V, & Kitajewski J.K. (2022). Notch1 and Notch4 core binding domain peptibodies exhibit distinct ligand-binding and anti-angiogenic properties. Angiogenesis, 26(2), 249-263.

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Immunohistochemical Detection of Tenascin-C in Lung

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To visualize TNC protein in the lung, paraffin-embedded lung sections were unmasked by boiling in a 10 mM pH 6 citrate solution. Slides were blocked with 5% normal goat serum in PBS and incubated with 1:100 primary anti-TNC antibody (AB19011, EMD Millipore, Temecula, CA). Slides were washed with 0.1% triton X-100 in PBS (Sigma-Aldrich, St Louis, MO) followed by 1:200 secondary anti-rabbit biotinylated antibody (Jackson ImmunoResearch, West Grove, PA). Slides were washed with 0.1% triton X-100 in PBS and treated with peroxide solution [0.6% H₂O₂ (Sigma-Aldrich, St Louis, MO) in methanol]. Slides were washed with 0.1% triton X-100 in PBS then were incubated with 3,3′-Diaminobenzidine developing solution (Vector Lab, Burlingame, CA) for 1 h at room temperature. The signal detection was done with the Elite ABC system (Vectastain, Burlingame, CA) and hematoxylin staining was performed. The sections were embedded into ProLong Gold antifade reagent (Invitrogen, Waltham, MA). Sections were examined using an Axio Imager A1. Pictures were taken with an AxioCam Icc3 camera and Axiovision software (all: Zeiss, Jena, Germany).

Meijer M.T., de Vos A.F., Scicluna B.P., Roelofs J.J., Abou Fayçal C., Orend G., Uhel F, & van der Poll T. (2021). Tenascin-C Deficiency Is Associated With Reduced Bacterial Outgrowth During Klebsiella pneumoniae-Evoked Pneumosepsis in Mice. Frontiers in Immunology, 12, 600979.

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Immunofluorescence Analysis of SCD-MSC and NS-MSC

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SCD-MSC and NS-MSC (2×10⁴ cell/well) grown on glass cover slips were fixed with 4% paraformaldehyde in PBS (pH 7.4) for 15 min at room temperature and permeabilized with 1% Triton X-100 (Sigma-Aldrich, Brazil). Cells were incubated with the following primary antibodies in 3% calf serum, 0.1%Triton X-100 in PBS (pH 7.4): rabbit anti-vimentin (1 : 200; Vector Laboratories) and mouse anti-SMA (1 : 200; Vector Laboratories). Cells were washed and incubated with secondary antibodies goat anti-rabbit Alexa Fluor 555 (Invitrogen) and goat anti-mouse Alexa Fluor 488 (Invitrogen) at 1 : 300 dilution. Nuclei were counterstained with DAPI (4,6-diamidino-2-phenylindole, 1 : 1000). Slides were examined with fluorescence confocal microscope (Leica TCS SP5 software; Leica Microsystems).

Ribeiro T.O., Daltro P.B., Daltro G.C., Freire S.M., Meyer R, & Fortuna V. (2020). Quantification and Comprehensive Analysis of Mesenchymal Stromal Cells in Bone Marrow Samples from Sickle Cell Disease Patients with Osteonecrosis. Stem Cells International, 2020, 8841191.

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