Escin, Con A, N-methyl-2-phenylindole, 5,5′-dithiobis (2-nitrobenzoic acid), and vanadium trichloride were supplied from Sigma (St Louis, MO, USA). N-(1-Naphthyl)-ethylenediaminedihydrochloride, carboxymethyl cellulose (CMC), Tris, acrylamide:bisacrylamide (29:1) 40% solution, sodium dodecyl sulfate, ammonium persulfate, and N,N,N′,N′-tetramethylethylenediamine were purchased from Fisher Chemical (Leicestershire, UK). Bovine serum albumen (BSA) and Tween 20 were purchased from MP Biomedicals (Irvine, CA, USA). All other chemicals were of the highest analytical grade available.
Tween 20
Manufactured by MP Biomedicals
Sourced in United States
Tween 20 is a non-ionic detergent commonly used in various laboratory applications. It is a polyoxyethylene sorbitan monolaurate, providing emulsifying, dispersing, and stabilizing properties. Tween 20 is widely utilized in scientific research and biochemical procedures due to its ability to solubilize and stabilize proteins, enzymes, and other biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Thermo Fisher Scientific (3 mentions)
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions)
Escin, by Merck Group (1 mentions)
Ammonium persulfate, by Thermo Fisher Scientific (1 mentions)
Vanadium trichloride, by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Thermo Fisher Scientific (1 mentions)
N methyl 2phenyl indole, by Merck Group (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
N 1 naphthyl ethylenediamine dihydrochloride, by Thermo Fisher Scientific (1 mentions)
N n n n tetramethylethylenediamine, by Thermo Fisher Scientific (1 mentions)
5 5 dithiobis 2 nitrobenzoic acid, by Merck Group (1 mentions)
2 2 azino bis, by Merck Group (1 mentions)
Hemin, by Merck Group (1 mentions)
Vimentin antibody, by Merck Group (1 mentions)
Neun antibody, by BioLegend (1 mentions)
Brightgreen 2x qpcr mastermix, by Applied Biological Materials (1 mentions)
Bovine serum albumin (bsa), by MP Biomedicals (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by MP Biomedicals (1 mentions)
Mops buffer, by Thermo Fisher Scientific (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
9 protocols using tween 20
1
Biochemical Assay Reagent Procurement
2
Hemin-Based Antioxidant Enzyme Assay
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Hemin purchased from Sigma Co. was used without further purification. 2,2ʹ′--azinobis(3--ethylbenzothiazoline--6--sulfonic acid) (ABTS) and bovine serum albumin (BSA), anti--FLAG antibody (monoclonal ANTI--FLAG, BioM2, produced in mouse, purified immunoglobulin) were obtained from Sigma Co. (St. Louis, MO, USA). 2--morpholinoethanesulfonic acid hydrate (MES) was purchased from Chemical Dojin Co. (Kumamoto, Japan); Triton X--100 detergent was obtained from Calbiochem Co. (Darmstadt, Germany), and Tween 20 was purchased from MP Biomedicals, Inc. (IIIkrich, France). Horseradish peroxidase (HRP), hydrogen peroxidase and other chemicals were obtained from Wako Pure Chemical Industries, Ltd. (Osaka, Japan).
3
Multilineage Cellular Characterization
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The reagents/materials used in the study are as follows: Bright green 2x qPCR Master Mix (Applied Biological Materials Inc., Canada), CD44 and CD45 antibodies (BD Biosciences, USA), NeuN antibody (Biolegend, USA), CD90 antibody (Cedarlane Cellutions Biosystems, Canada), glial fibrillary acidic protein(GFAP)antibody (Cloud Clone, USA), bovine serum albumin, DAPI, Tween-20 (MP Biomedicals, USA), β-III tubulin antibody (R&D Systems, USA), Alexa fluor 546 rat anti-rabbit secondary antibody (Santa Cruz Biotechnology Inc., USA), AP, MTT, TQ, Triton X-100, Vimentin antibody (Sigma Aldrich, USA), c-kit (CD117) antibody, DMEM, L-glutamine, penicillin-streptomycin, RevertAid First Strand cDNA synthesis kit, Sodium pyruvate, Trypsin EDTA (Thermo Fisher Scientific, USA),
4
Western Blot of Signaling Pathways
5
PFOA Exposure in Mice
6
Bacterial Biomass Determination in Seawater
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For the L–L assay, a commercially available kit for bacterial biomass determination (CheckLite HS Set, Kikkoman Biochemifa Co., Tokyo, Japan) containing the ATP releasing (cell lysis) reagent and the L–L reagent was modified for seawater sample measurement [18 ,19 (link)]. EDTA (ethylenediaminetetraacetic acid, Wako Pure Chemical Industries, Ltd., Osaka, Japan) was added to the ATP releasing reagent at a final concentration of 10 mM to avoid precipitation in the presence of the seawater samples or the seawater-based ATP standard solutions. To avoid the adsorption of the reagents and the adhesion of natural particles or debris to the micro-channels, 2% (v/v) Tween 20 (MP Biomedicals LLC, Santa Ana, CA, USA) was added to the L–L reagent. Both EDTA and Tween 20 were sterilized by autoclaving prior to use to eliminate potentially contaminating ATP. Seawater-based ATP standard solutions (5 × 10−12, 5 × 10−11, 5 × 10−10 M ATP and blank) were prepared from an original ATP standard solution (2 × 10−6 M ATP, Kikkoman Biochemifa Co., Tokyo, Japan) by diluting it with autoclaved artificial seawater (Daigo’s artificial seawater SP for marine microalgae medium, Nihon Pharmaceutical Co., Ltd., Tokyo, Japan). All reagents and standards were aseptically introduced into sterilized plastic bags (DSF-300, Tsukada Medical Research Co. Ltd., Ueda, Japan) for use in the in-situ analyzer.
7
Serological Detection of GLRaV-3 Virus
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The present study used a mixture of MAb and PAb specific to GLRaV-3 (Bioreba, Reinach, Switzerland); GAMI and GARI (Arista Biologicals, Allentown, PA, USA); streptavidin–horseradish peroxidase conjugate, 3,3’,5,5’-tetramethylbenzidine dihydrochloride (TMB), polyvinylpyrrolidon K25 (PVP) with m.v. ~24 kDa, polyethylenglycol (PEG) with m.w. ~6 kDa, Tris, sucrose, Triton X-100, sodium citrate, chloroauric acid, sodium azide, agarose (Sigma-Aldrich, St. Louis, MO, USA), dimethyl sulfoxide (DMSO), Tween 20 (MP Biomedicals, Santa Ana, CA, USA), and bovine serum albumin (BSA) (Biowest, Nuaillé, France). All auxiliary reagents (acids, alkalis, salts, and organic solvents) were of analytical or chemical purity grade.
Solutions for syntheses of the GNPs and their conjugates with antibodies were prepared using deionized water, the resistivity of which at 25 °C was no less than 18.2 MΩ·cm (Simplicity, Millipore, Burlington, MS, USA).
ELISA was conducted using 96-well transparent microtitration plates (Costar 9018, Corning Costar, New York, NY, USA). To manufacture test strips, the following membranes were used: a nitrocellulose (NC) membrane grade CNPC with a pore size of 15 μm attached to a solid support, a conjugate release matrix PT-R7, a separation membrane GFB-R4, and an absorption membrane AP045 (all membranes are from Advanced Microdevices, Ambala Cantt, India).
Solutions for syntheses of the GNPs and their conjugates with antibodies were prepared using deionized water, the resistivity of which at 25 °C was no less than 18.2 MΩ·cm (Simplicity, Millipore, Burlington, MS, USA).
ELISA was conducted using 96-well transparent microtitration plates (Costar 9018, Corning Costar, New York, NY, USA). To manufacture test strips, the following membranes were used: a nitrocellulose (NC) membrane grade CNPC with a pore size of 15 μm attached to a solid support, a conjugate release matrix PT-R7, a separation membrane GFB-R4, and an absorption membrane AP045 (all membranes are from Advanced Microdevices, Ambala Cantt, India).
8
Immunocytochemical Analysis of iPSCs
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For whole-mount immunocytochemical analysis, iPS cells cultures were fixed in 4 % paraformaldehyde (FUJIFILM Wako Pure Chemical Corporation) for 30 min, followed by permeabilization with 0.1 % Triton-X (nakalai tesque) in phosphate-buffered saline (PBS; Thermo Fisher Scientific) for 10 min at room temperature, washed once with 0.1 % Tween-20 (MP, Irvine, CA, USA) in PBS (PBST) then incubated with 20 % Blocking One (nakalai tesque) in PBS-T in a humidified chamber for 1 h at room temperature. The cells were incubated with diluted (1:200) antibody in 20 % Blocking One in PBS-T. After washing 3 times with PBS-T, the cells incubated with diluted (1:1000) secondary antibody in 20 % Blocking One for 2 h at room temperature in the dark. After washing off the secondary antibody in PBS-T, cells were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). The following antibodies were used as first antibodies: Mouse anti-OCT3/4 antibody (Santa Cruz Biotechnology, Dallas, TX, USA), goat anti-SOX17 antibody (R&D systems), Goat anti-human Albumin antibody (Fortis Life Sciences, Montgomery, TX, USA). Secondary antibodies used were Alexa 488-conjugated and Alexa 568-conjugated antibodies (Thermo Fisher Scientific).
9
Cornea Immunofluorescence Staining Procedure
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After incubation, cornea quarters were fixed in methanol chilled to −20 for 30 min, rinsed at room temperature in PBS containing 0.05% Tween-20 (Sigma), and blocked for 30 min at 37°C in PBS containing 2% bovine serum albumin (Fisher Scientific, Pittsburgh, PA) and 2% goat serum (Sigma, St. Louis, MO) (blocking solution). Quarters were subsequently incubated in 2.5 μg/mL mouse anti-ZO1 (clone 1A12; Thermo Scientific) in the above blocking solution for 60 min at 37°C, rinsed 3 times in PBS containing 0.05% Tween-20, and fixed at room temperature for 30 min in 0.5% paraformaldehyde (Electron Microscopy Sciences, Hatfield, PA) in PBS. The quarters were then rinsed again in PBS containing 0.05% Tween-20 and permeabilized at room temperature for 5 min in PBS containing 1% Triton X-100 (MP Biomedicals, Santa Ana, CA) followed by 3 rinses of 5 min each in PBS.
The EdU click-it reaction with Alexa Fluor 488 and subsequent staining with Hoechst 33342 was performed as described previously.37 (link) Next, quarters were rinsed again in PBS containing 0.05% Tween-20 for 30 min and incubated in blocking solution containing Alexa Fluor 555-labeled goat anti-mouse IgG (Thermo Scientific) for 60 min at 37°C and rinsed again 3 times in PBS containing 0.05% Tween-20 for 30 min.
The EdU click-it reaction with Alexa Fluor 488 and subsequent staining with Hoechst 33342 was performed as described previously.37 (link) Next, quarters were rinsed again in PBS containing 0.05% Tween-20 for 30 min and incubated in blocking solution containing Alexa Fluor 555-labeled goat anti-mouse IgG (Thermo Scientific) for 60 min at 37°C and rinsed again 3 times in PBS containing 0.05% Tween-20 for 30 min.
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