LPL activity was determined by measuring the generation of non-esterified fatty acids (NEFA). In a 96-well plate, 0–2 µM of an ApoC2 mimetic peptide, 0.83 nM of LPL from bovine milk (Sigma-Aldrich), ApoC2-deficient ethylenediaminetetraacetic acid (EDTA)-plasma (diluted with PBS to a final TG concentration of 0.67 mg/dl), 2 IU/ml heparin, 0.2% fatty acids free bovine serum albumin (BSA) (ICN Biomedicals), and PBS (Life Technologies) were combined in a final reaction volume of 50 μl. The reaction mixture was pre-incubated on ice for 30 min and incubated at 37°C for 1 h. NEFA were measured with a coupled enzyme reaction (Wako Diagnostics) in a Synergy H1 microplate reader (BioTek, USA).
Phosphate buffered saline (pbs)
Manufactured by ICN Biomedicals
Sourced in United States
Phosphate-Buffered Saline (PBS) is a commonly used buffer solution designed to maintain the pH and osmolarity of biological samples. It is a widely used reagent in various laboratory applications, including cell culture, immunoassays, and biochemical studies. PBS is a simple, isotonic, and pH-balanced solution that mimics the physiological environment, making it a versatile and essential tool for researchers and scientists.
Automatically generated - may contain errors
Lab products found in correlation
Bovine milk, by Merck Group (1 mentions)
Synergy h1 microplate reader, by Agilent Technologies (1 mentions)
Bovine serum albumin bsa, by ICN Biomedicals (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Dmem low glucose medium, by Merck Group (1 mentions)
T75 flask, by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Glutaraldehyde aqueous solution, by Merck Group (1 mentions)
Aptes, by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
3 protocols using phosphate buffered saline (pbs)
1
Lipoprotein Lipase Activity Assay
2
Isolating and Irradiating hAM-MSCs
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Human placentas were collected during caesarean section births of healthy women after approval of the local ethics board and obtaining informed consent. Samples of AM were collected in sterile conditions by a specialist at Milad Hospital in Tehran and transferred in a tube containing phosphate-buffered saline (PBS, ICN Biomedicals, Ohio, USA) and antibiotics. Under sterile condition, the AM was isolated from placenta and cut into small pieces. The pieces were washed several times with PBS containing antibiotics (Cinagen, Karaj, Iran) and digested with collagenase type I (Gibco, Grand Island, USA) in a shaking incubator at 37 °C for 1–2 hr. To clear cell suspensions of unwanted materials, they were filtered and centrifuged at 750 g for 5 min. The cells were introduced into the DMEMLow Glucose medium (Sigma Aldrich, Gillingham, Wisconsin, USA) contained FBS (Invitrogen, Camarillo, California, USA) at a final concentration of 10%, Penicillin-Streptomycin (10,000 units penicillin and 10 mg streptomycin/ml) and were seeded into T75 flasks (Nunc, Roskilde, Denmark). The flasks were placed in an incubator at 37 °C in humidified atmosphere with 5% CO
2
. hAM-MSCs at passage 3 were irradiated with gamma rays from a Cs-137 source (30 Gy or 3000 rad) and used for co-culture with T-cells.
2
. hAM-MSCs at passage 3 were irradiated with gamma rays from a Cs-137 source (30 Gy or 3000 rad) and used for co-culture with T-cells.
3
Protein Immobilization on Mica Surfaces
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Other reagents used in the experiments were: (a) phosphate buffered saline (PBS, ICN Biomedicals, pH 7.4, containing 10 mM of PO42−, 137 mM of NaCl and 27 mM of KCl) was used to prepare all protein solutions; (b) 3-aminopropyltriethoxysilane (APTES, Sigma) was used for the silanization of the mica and cantilever surfaces; (c) 2.5% glutaraldehyde aqueous solution, prepared from a 25% solution of glutaraldehyde was purchased from Sigma. All solutions were prepared using deionized water (Cobrabid water purification system, 0.08 µS).
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