Transgenic CL2006 worms synchronized to the L4 stage (early adult stage) were inoculated on NGM plates with or without drugs, and N2 was used as a negative control for Aβ deposition (32 (link)). Following incubation at 20˚C for 48 h, the worms were collected with M9 buffer and fixed in 4% paraformaldehyde/PBS (pH 7.4) (cat. no. BL539A; Biosharp Life Sciences) at 4˚C for 24 h. The worms were then treated with 5% β-mercaptoethanol (cat. no. M828395; Macklin, Inc.), 1% Triton X-100 (cat. no. MB2486; Dalian Meilun Biology Technology Co., Ltd.) and 125 mM Tris (pH 7.4) (cat. no. T8060; Beijing Solarbio Science & Technology Co., Ltd.) at 37˚C for 24 h. The samples were stained with 0.125% thioflavin S (cat. no. S19293; Shanghai Yuanye Bio-Technology Co., Ltd.) in 50% ethanol at room temperature for 2 min, and then with 50% ethanol. The samples were thereafter rinsed and transferred to a glass slide for observation under a laser scanning confocal microscope (LSM900; Carl Zeiss AG). The number of thioflavin S-reactive deposits in the area anterior to the pharyngeal bulb was counted.
Triton x 100
Manufactured by Maclin Biochemical Technology
Sourced in China
Triton X-100 is a non-ionic detergent commonly used as a laboratory reagent. It is a surfactant that can be used for cell lysis, protein extraction, and various other applications in biochemical and cell biology research.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Lsm 900, by Zeiss (2 mentions)
Thioflavin s, by Solarbio (2 mentions)
β mercaptoethanol, by Biosharp (2 mentions)
Dm6000b fluorescence microscope, by Leica camera (1 mentions)
Alexa fluor 488 or 594 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Application suite x software, by Leica camera (1 mentions)
Goat serum, by Beyotime (1 mentions)
Disc3 5, by Maclin Biochemical Technology (1 mentions)
Phosphate buffered saline solution, by Maclin Biochemical Technology (1 mentions)
Polymyxin b, by Maclin Biochemical Technology (1 mentions)
Transdetect cell counting kit 8 cck 8, by Transgene (1 mentions)
Ultrapure water machine, by Merck Group (1 mentions)
Caspase 3 19677 1 ap, by Proteintech (1 mentions)
Survivin sirna, by GenePharma (1 mentions)
β actin 20536 1 ap, by Proteintech (1 mentions)
Huvec cells, by American Type Culture Collection (1 mentions)
Coralite 594 conjugated secondary antibodies, by Proteintech (1 mentions)
Horseradish peroxidase conjugated, by Proteintech (1 mentions)
15 protocols using triton x 100
1
Quantifying Amyloid-beta Deposition in Transgenic Worms
2
Immunostaining of Pancreatic and Cell Samples
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Paraffin-embedded sections of the pancreas of rats were deparaffinized and rehydrated, followed by antigen retrieval in 10 mmol/L sodium citrate buffer. After blocking in 5% goat serum (Beyotime) for 30 min, the sections were incubated with antibodies against insulin (no. 66198-1-Ig, ProteinTech), Nrf2 (no. 16396-1-AP, ProteinTech), HO-1 (no. 10701-1-AP, ProteinTech), and Grx1 (no. A5315, ABclonal, Wuhan, China) at 4 °C overnight, followed by incubation with Alexa Fluor 488- or 594-conjugated secondary antibodies (1:400 dilution, Molecular Probes, Waltham, MA, USA). DAPI was used for nuclei staining.
For immunostaining of cultured RIN-m5F cells, the cells were fixed in 4% formaldehyde for 15 min and then incubated in 0.5% Triton-X100 (MACKLIN, Shanghai, China) for 15 min. After blocking with 3% bovine serum albumin at room temperature for 1 h, the cells were incubated overnight at 4 °C with an Nrf2 antibody. This was followed by a 1-h incubation with an Alexa Fluor 488-conjugated secondary antibody (1:500 dilution, Molecular Probes) after washing. Negative control sections were treated with PBS instead of a primary antibody. The antibodies were visualized using a Leica DM6000B fluorescence microscope (Wetzlar, Germany) with the Leica Application Suite X software (3.6.1.23246).
For immunostaining of cultured RIN-m5F cells, the cells were fixed in 4% formaldehyde for 15 min and then incubated in 0.5% Triton-X100 (MACKLIN, Shanghai, China) for 15 min. After blocking with 3% bovine serum albumin at room temperature for 1 h, the cells were incubated overnight at 4 °C with an Nrf2 antibody. This was followed by a 1-h incubation with an Alexa Fluor 488-conjugated secondary antibody (1:500 dilution, Molecular Probes) after washing. Negative control sections were treated with PBS instead of a primary antibody. The antibodies were visualized using a Leica DM6000B fluorescence microscope (Wetzlar, Germany) with the Leica Application Suite X software (3.6.1.23246).
3
PEDOT:PSS-Wrapped MIL-101(Cr)/S Composites
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Poly(3,4-ethylenedioxythiophene)/poly(styrene sulfonate) (PEDOT:PSS) solution was prepared by filtering commercially available solution (∼1 wt% solid content, Clevios PH1000) and adding 5 wt% dimethyl sulfoxide (DMSO), and 1 wt% Triton X-100 (biotechnology grade, MACKLIN, China) (final PEDOT:PSS concentration is ∼0.94 wt%).
Conductive-polymer solution and MIL-101(Cr)/S were mixed with a mass ratio of 1 : 10, and then the homogeneous mixture is milled in an agate mortar for about half an hour to reduce the particle sizes. The resulting powder was dried at 120 °C to remove water in the conductive-polymer solution, and the process is repeated several times till MIL-101(Cr)/S is fully wrapped in scale-like structures.
Conductive-polymer solution and MIL-101(Cr)/S were mixed with a mass ratio of 1 : 10, and then the homogeneous mixture is milled in an agate mortar for about half an hour to reduce the particle sizes. The resulting powder was dried at 120 °C to remove water in the conductive-polymer solution, and the process is repeated several times till MIL-101(Cr)/S is fully wrapped in scale-like structures.
4
Bacterial Membrane Permeability Analysis
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Mueller–Hinton broth (MHB) and Mueller–Hinton agar (MHA) were purchased from GL Biochem (Shanghai, China). Triton X-100 and phosphate-buffered saline (PBS) solution were purchased from Macklin (Shanghai, China). BCECF-AM, EtBr, PI, diSC3(5), polymyxin B, 4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid (HEPES) and NPN were also purchased from Macklin (Shanghai, China). TransDetect Cell Counting Kit-8 (CCK-8) and Dulbecco’s modified Eagle’s medium with high glucose (DMEM) were purchased from TransGen Biotech (Beijing, China). Fetal bovine serum (FBS) was purchased from Gibco (Shanghai, China).
5
Survivin siRNA Transfection in Glioma Cells
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All oligonucleotides were synthesized and purified by Sangon Biotech Company (Shanghai, China). Survivin siRNA (5ʹ-AUUCACCAAGGGUUAAUUCdTdT-3ʹ) was synthesized by GenePharma Company (Shanghai, China). Deionized water used in all aqueous solutions was produced using a Millipore Ultrapure water machine (Massachusetts, USA). GelRed DNA gel stain solution was purchased from Biosharp (Beijing, China). Tris, magnesium chloride (MgCl2), and Triton X-100 were obtained from Macklin (Shanghai, China). Agarose was purchased from Biowest (Barcelona, Spain). U87, U251, and HUVEC cells were purchased from ATCC. Fetal bovine serum (FBS) and high-glucose Dulbecco’s modified Eagle’s medium (DMEM/high glucose) were bought from Gibco (NY, USA). A cell counting kit (CCK-8) was purchased from Bimake (Texas, USA). 4′6-Diamidino-2-phenylindole (DAPI) were obtained from Solarbio (Beijing, China). Primary antibodies β-actin (20536-1-AP), NCL (10556-1-AP), and Caspase-3 (19677-1-AP) were obtained from Proteintech (IL, USA). Survivin (YT4472) was purchased from Immunoway (TX, USA), and horseradish peroxidase-conjugated or CoraLite594-conjugated secondary antibodies were purchased from Proteintech (IL, USA).
6
Quantifying Amyloid Deposition in Transgenic C. elegans
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Transgenic C. elegans strain CL2006 synchronized to the L4 stage (early adult stage) was inoculated on NGM plates with or without EEGE. N2 worms were used as a negative control for Aβ deposition (28 (link)). After incubation at 20˚C for 48 h, worms were collected with M9 buffer and fixed in 4% paraformaldehyde/PBS (pH 7.4) (cat. no. BL539A; Biosharp Life Sciences) at 4˚C for 24 h. The worms were then incubated in 5% β-mercaptoethanol (cat. no. M828395; Macklin, Inc.), 1% Triton X-100 (cat. no. MB2486; meilunbio) and 125 mM Tris (pH 7.4) (cat. no. T8060; Beijing Solarbio Science & Technology Co., Ltd.) at 37˚C for 24 h. The worms were then stained with 0.125% thioflavin S (cat. no. S19293; Shanghai Yuanye Biotechnology Co., Ltd.) in 50% ethanol at room temperature for 2 min, and then rinsed in 50% ethanol 2-3 times. The worms were then placed on a glass slide for observation under a laser scanning confocal microscope (LSM900; Carl Zeiss AG). The amount of thioflavin S deposition in the prepharyngeal region of each nematode was scored to quantify amyloid deposits.
7
Cytotoxicity Evaluation of Nanomaterials
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Paclitaxel, SA, MIF, DXM, pyridine, Na2CO3, paraformaldehyde, FITC, DAPI, Triton X‐100, MTT, and D‐luciferin potassium salt were all purchased from Shanghai Macklin Biochemical Co., Ltd. All the solvents, including acetone, methanol, polyoxyethylene castor oil, ethanol, and DMSO were obtained from J&K Scientific Ltd, and used without further purification. DMEM, RPMI‐1640, FBS, and penicillin‐streptomycin solution were purchased from Gibco Life Technologies. MCF‐7, MDA‐MB‐231, and 4T1 cells were maintained in our laboratory.
8
Enzymatic Assay for Oxidative Stress
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Lysozyme, glutathione (GSSG), dithiothreitol (DTT), H2O2, 2,6-dimethoxyphenol (DMP), 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), phenylmethanesulfonyl fluoride (PMSF), and isopropyl-β-D-thiogalactopyranoside (IPTG) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Triton X-100, Tween-80, ethanol, and other chemicals were obtained from Macklin (Shanghai, China) and were of analytical grade.
9
Squid Protein Extraction and Characterization
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Glycine, acrylamide solutions with 30% (w/v), SDS, ethylene diamine tetraacetic acid (EDTA), dithio-bis-nitrobenzoic acid (DTNB), 1-anilino-8-naphtha-lene sulfonate (ANS), Triton-X100, N,N,N′,N′-tetramethylethylenediamine (TEMED), and ammonium persulfate (APS) were provided by Macklin Biochemical Co., Ltd. (Shanghai, China). Ready-to-use protein molecular weight standard marker (6.5–200 kDa) was purchased from Sigma (St. Louis, MO). Disodium hydrogen phosphate (Na2HPO4), potassium dihydrogen phosphate (KH2PO4), potassium chloride, sodium hydroxide, ethanol, and other reagents were obtained from Hangzhou Jiecheng Biotechnology Co. (Hangzhou, China). All reagents were of analytical purity.
Giant squid, purchased from Zhoushan Second Ocean Fishery Co., Ltd. (Zhoushan, China), were immediately frozen without any treatment after being caught and stored at −20°C for less than 3 months. All squids were deheaded, degutted, and cleaned using water after they were defrosted in the air to room temperature. The mantles with the fins were sealed and stored at −40°C for further experiments.
Giant squid, purchased from Zhoushan Second Ocean Fishery Co., Ltd. (Zhoushan, China), were immediately frozen without any treatment after being caught and stored at −20°C for less than 3 months. All squids were deheaded, degutted, and cleaned using water after they were defrosted in the air to room temperature. The mantles with the fins were sealed and stored at −40°C for further experiments.
10
Fabrication of Potentiometric Ion Sensors
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CB was purchased from Cabot Corporation. Graphite was obtained from Qingdao Jintao Graphite Co., Ltd. Defoamer (BYK-022) was purchased from BYK Chemistry. Dispersant (T-859) and waterborne polyurethane resin were purchased from Shenzhen Jitian Chemical Co., Ltd. PEDOT/PSS was obtained from 3AMaterial. Sodium ionophore III (ETH2120) was purchased from Shanghai Yuanye Biotechnology Co., Ltd (MedMol). Potassium tetrakis-(4-chlorophenyl)-borate (KTClPB) and bis-(2-ethylhexyl)-sebacate (DOS) was purchased from Aladdin. Polyvinyl chloride (PVC), cyclohexanone (CHA), and TritonX-100 were got from Shanghai Macklin Biochemical Co., Ltd. Ag/AgCl paste was purchased from Shanghai Julong Electronic Technology Co., Ltd. All chemicals and solvents were used as received without further treatment.
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