BHK-21 cells were seeded in 12-well plates, and after 24 h of culture, they were rinsed with phosphate-buffered saline (PBS) (Solarbio, Beijing, China) and fixed using methanol (Solarbio) for 10 min at room temperature. The cells were washed three times with PBS and then incubated in blocking buffer [3% bovine serum albumin (BSA) (BOSTER, Beijing, China) in Tris-buffered saline (Solarbio) and Tween20 (Solarbio)] for 1 h at room temperature. The cells were then incubated with an anti-E monoclonal antibody (Abcam, Cambridge, UK) (1:20) in PBS with 3% BSA overnight at 4°C. After the cells were washed three times in PBS, they were incubated with a fluorescently labeled secondary antibody—fluorescein isothiocyanate-conjugated goat antimouse antibody (ZSGB-Bio, Beijing, China)—in the dark at 37°C for 1 h. The nuclei were washed with PBS three times and stained with 4,6-diamidino-2-phenylindole (DAPI) (Solarbio). Slides were imaged under a fluorescence microscope (Olympus, Tokyo, Japan).
Methanol
Manufactured by Solarbio
Sourced in China
Methanol is a colorless, volatile, and flammable liquid. It is a versatile chemical compound with a wide range of applications in various industries, including as a fuel, solvent, and chemical feedstock. Methanol is commonly used in laboratories for various purposes, such as cleaning and extraction procedures.
Automatically generated - may contain errors
Lab products found in correlation
Crystal violet, by Solarbio (10 mentions)
Phosphate buffered saline (pbs), by Solarbio (5 mentions)
Crystal violet, by Merck Group (4 mentions)
Matrigel, by Corning (3 mentions)
Inverted microscope, by Olympus (2 mentions)
A microscope, by Olympus (2 mentions)
Matrigel, by BD (2 mentions)
Anti e monoclonal antibody, by Abcam (1 mentions)
Bovine serum albumin (bsa), by Abcam (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Tween 20, by Solarbio (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Wright s stain, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Lutein, by Solarbio (1 mentions)
Proelut c18, by Dikma Technologies (1 mentions)
Proelut psa, by Dikma Technologies (1 mentions)
Proelut carb, by Dikma Technologies (1 mentions)
Potassium nitrate, by Sinopharm (1 mentions)
25 protocols using methanol
1
Immunofluorescence Staining of BHK-21 Cells
2
Antileishmanial Activity of Sodium Stibogluconate
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RAW 264.7 cells (~ 5.0 × 105 cells/per well) were plated on round glass coverslips in 24-well plates and allowed to adhere to the slides for 24 h at 37 °C, 5% CO2 with RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% FBS (Gibco). Stationary phase promastigotes (~ 5.0 × 106 cells/per well) of HCZ isolate, DD8 and 9044 strains were added into wells, and incubated at 37 °C, 5% CO2 for 6 h. Next, the non-infected promastigotes were removed, and the infected macrophages were incubated at 37 °C in 5% CO2 with fresh medium without drugs for 24 h. The medium was then removed, and fresh culture medium supplemented with 10 μM SSG (cf. [33 (link)]) were added into wells for 2 days, with PBS used as control. At 24 and 48 h, the glass coverslips were fixed in methanol (Solarbio, Beijing, China) and stained with Wright’s stain (Solarbio) to calculate the number of intracellular amastigotes under light microscopy by counting 200 macrophages per slide. Inhibition rate (%) = 100% − [(No. of amastigotes in treated sample/ No. of amastigotes in control) × 100%].
3
Extraction and Analysis of Lutein
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All chemicals
were of analytical grade unless otherwise indicated. Potassium nitrate
(purity ≥99.0%) was purchased from Sinopharm (Beijing, China).
Lutein (HPLC ≥90%) and methanol (HPLC grade) were purchased
from Solarbio (Beijing, China) and Fisher Chemicals (Loughborough,
UK). Ultrapure water was produced by a Milli-Q purification system.
The SPE cartridge ProElut AC 250 mg/3 mL, ProElut C18 500 mg/3 mL,
ProElut CARB 250 mg/3 mL, ProElut CARB 500 mg/6 mL, ProElut NH2 500 mg/3 mL, ProElut PSA 500 mg/3 mL, ProElut CARB/NH2 500/500 mg/6 mL and ProElut CARB/PSA 500/500 mg/6 mL were
purchased from Dikma (Beijing, China).
were of analytical grade unless otherwise indicated. Potassium nitrate
(purity ≥99.0%) was purchased from Sinopharm (Beijing, China).
Lutein (HPLC ≥90%) and methanol (HPLC grade) were purchased
from Solarbio (Beijing, China) and Fisher Chemicals (Loughborough,
UK). Ultrapure water was produced by a Milli-Q purification system.
The SPE cartridge ProElut AC 250 mg/3 mL, ProElut C18 500 mg/3 mL,
ProElut CARB 250 mg/3 mL, ProElut CARB 500 mg/6 mL, ProElut NH2 500 mg/3 mL, ProElut PSA 500 mg/3 mL, ProElut CARB/NH2 500/500 mg/6 mL and ProElut CARB/PSA 500/500 mg/6 mL were
purchased from Dikma (Beijing, China).
4
Bioactive Compound Extraction from Hemerocallis citrina
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Hemerocallis citrina Borani was obtained from Qingyang, Gansu, China. Standards of glucose, mannose, fucose, rhamnose, xylose, galactose, arabinose, glucuronic acid, ribose, and galacturonic acid were purchased from Shanghai Yuanye Biotechnology (Shanghai, China). Absolute ethanol was purchased from Fuyu Fine Chemical (Tianjin, China). Trifluoroacetic acid, methanol, sodium hydroxide, hydrochloric acid, carbazole, 1-phenyl-3-methyl-5-pyrazolone, 3,5-dinitrosalicylic acid, sulfuric acid, Coomash bright blue, chloroform, potassium bromide, sodium azide, sodium acetate, anthranone, and other reagents were purchased from Solarbio Biotechnology (Beijing, China). ABTS and FRAP assay kits were purchased from Suzhou Keming Biotechnology (Suzhou, China). Distilled water was used in these experiments.
5
Colony Forming Assay Protocol
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1 × 103 cells were cultured in 6-well plates for 2 weeks. After fixed in methanol (Solarbio, Beijing, China), colonies were processed with crystal violet (Sigma-Aldrich). Visible colonies were counted manually under microscope (Olympus, Tokyo, Japan).
6
Anlotinib and Metformin Cytotoxicity Assay
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Anlotinib was kindly provided by Chia Tai Tian Qing Pharmaceutical Group Co., Ltd (Nanjing, China). Metformin, 3- (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), methanol, crystal violet, and phosphate-buffered saline (PBS) were purchased from Solarbio Bioscience & Technology Co. Ltd (Beijing, China). Hoechst 33342, propidium iodide (PI), 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA), and SDS lysis buffer were obtained from Beyotime Biotechnology Co., Ltd (Shanghai, China). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), trypsin, and penicillin/streptomycin were purchased from Invitrogen (Carlsbad, CA, United States).
7
In Vitro Cell Invasion Assay
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T24 and HT-1197 cells in apical compartment were cultured with serum-free medium, and the basolateral chamber was filled with 10% FBS medium. Cells were plated on the top compartment of membrane pre-coated with Matrigel for invasion assay. 24 h later, invaded cells were fixed in methanol (Solarbio, Beijing, China) and dyed in crystal violet (Solarbio). The cell numbers were recorded and photographed randomly from 5 chosen fields by an inverted microscope (Olympus, Tokyo, Japan).
8
GC Cell Fixation and Staining
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After transfection, GC cells were maintained in 6-well plates and incubated for 2 weeks. Later, cells in each well were fixed and dyed by methanol (Solarbio, Beijing, China) or crystal violet (Solarbio) for 10 min or 5 min, respectively.
9
Transwell Assay for Cell Invasion and Migration
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Transwell chamber which was coated with or without Matrigel (BD Biosciences, Massachusetts, USA) was applied to measure cell invasion or migration, respectively. MKN-45 or HGC-27 cells in serum-free media were placed into the top chamber. Then, media with 10% FBS was added into the basolateral chamber. 48 h later, invaded or migrated cells were fixed in methanol (Solarbio), stained with 0.1% crystal violet (Solarbio) and counted via a microscope (Olympus).
10
Colony Formation Assay Protocol
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Culture of the logarithmic cells of 1 × 103 was in 6-well plates. After fixation in methanol (Solarbio, Beijing, China), treatment of the colonies was with crystal violet (Sigma-Aldrich). Manual counting of visible colonies was performed under a microscope (Olympus, Tokyo, Japan) [17 (link)].
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