Antibodies against Nrf2 (Cat# 12712), NF-κB p65 (Cat# 3034), collagen A1 (Cat# 66948), collagen 3A (Cat#30565), alpha-smooth muscle actin (α-SMA) (Cat#19245), β-actin (Cat# 4970), and Lamin A (Cat# 86846) were purchased from Cell Signalling, USA. Antibodies against TGFβ-1 (Cat# sc-130348), smad3 (Cat# sc-101154), smad7 (Cat# sc-365846), phospho-smad3 (Ser425) (Cat# sc-517575), and BMP7 (Cat# sc-53917) were purchased from Santa Cruze biotechnology, USA. A Bradford-based protein assay kit (Cat# 23200), phosphate buffer saline solution (Cat# 20012043), protease inhibitor cocktail (Cat# A32965), radio-immuno-precipitation assay RIPA buffer (RIPA) buffer (Cat# ab156-34); and nuclear-cytoplasmic/nuclear extraction kit (Cat# 78833) were purchased from ThermoFisher scientific.
Phosphate buffer saline solution
Manufactured by Thermo Fisher Scientific
Sourced in United States
Phosphate buffer saline solution is a common buffer solution used in various laboratory applications. It is an aqueous solution containing sodium phosphate and sodium chloride, which helps maintain the pH and osmolarity of the sample. The solution is typically used to dilute, stabilize, and preserve biological samples, such as proteins, cells, and tissues, during experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor cocktail, by Thermo Fisher Scientific (2 mentions)
Antibodies against nrf2, by Cell Signaling Technology (1 mentions)
Lamin a, by Cell Signaling Technology (1 mentions)
Alpha smooth muscle actin α sma, by Cell Signaling Technology (1 mentions)
Nf κb p65, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Agarose, by Merck Group (1 mentions)
Trypsin edta 1x, by Thermo Fisher Scientific (1 mentions)
Direct q water system, by Merck Group (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
Wst 8, by Abcam (1 mentions)
Igf 1, by Thermo Fisher Scientific (1 mentions)
Fluorescein isothiocyanate fitc phalloidin, by Merck Group (1 mentions)
Antibiotic antimycotic solution, by Thermo Fisher Scientific (1 mentions)
Chloroform, by Merck Group (1 mentions)
4 protocols using phosphate buffer saline solution
1
Western Blot Analysis of Fibrosis Markers
2
High-Throughput 3D Glioblastoma Spheroid Assay
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Spheroids were produced using a variation of the liquid-overlay 96-well plate method 52 . A solution of 1% (v/w) agarose (Sigma Aldrich) was prepared in deionized water and autoclaved. After heating up the solution one minute in the microwave, 75 µL was added in each well to form a meniscus on the bottom of the well. After 10 minutes at room temperature, the agarose cooled down and jellified to form a nonadherent round-bottom well. Glioblastoma cells were rinsed with Phosphate Buffer Saline solution (Thermofisher D8537) and detached using 1 mL of 0.25% Trypsin-EDTA (1X) (Gibco 25200-056) at 37 °C. The desired quantity of cells was added per well, in a total volume of 150 µL per well. Half of the media was changed every 2-3 days. This method of generating spheroids is reproducible, inexpensive and high-throughput. Indeed, it requires 7 days and only very little material to produce hundreds of spheroids with a similar size exceeding 400 µm. Moreover, spheroids are easy to handle and can be collected and grafted using a standard 1000-µL pipette.
3
Cell Culture Reagent Preparation
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All culture media and reagents were purchased from Sigma Chemical Company (St. Louis, MO, USA), except where otherwise indicated. Phosphate buffer saline (PBS) solution was supplied by Gibco/BRL (Grand Island, NY, USA). Methanol (ACS/HPLC) grade was purchased from Burdik & Jackson (Muskegon, MI, USA). Ultrapure water, purified by Direct-Q water system (Millipore, Bedford, MA, USA), was used for the preparation of solvents.
4
Skeletal Muscle Cell Culture Protocol
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2.1. Materials and methods PCL of mol. wt. ∼90 000, chloroform, acetic acid and N, N-dimethyleformamide (DMF) were purchased from Merck, Germany. Skeletal muscle growth media and skeletal muscle differentiation media (Promocell, Germany); insulin like growth factor 1 (IGF-1) (Invitrogen, USA), fetal bovine serum (FBS), horse serum, antibiotic-antimycotic solution, phosphate buffer saline (PBS) solution (GIBCO, USA); paraformaldehyde, dimethyl sulfoxide (DMSO) (Sigma Aldrich, USA); all primary and secondary antibodies (Abcam, United Kingdom);WST-8 [2-(2-methoxy-4nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt], collagen type-1 (rat tail) and fluorescein isothiocyanate (FITC)-phalloidin (Sigma Aldrich, USA) were purchased and used as received.
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