Blood samples were centrifuged at 5000 rpm for 5 mins to separate the plasma. Measured volumes (not exceeding 20 µL) of plasma, CSF, and injectate were transferred into scintillation tubes. 5 mL of scintillation fluid (Emulsifier-safe, PerkinElmer) was then added. Tissue samples were weighed and up to 50 mg were dissolved in 500 µL of Soluene 350 (PerkinElmer) overnight in a scintillation tube. Two drops of acetic acid were added to each tube to neutralise the alkaline Soluene 350, followed by 5 mL of scintillation fluid (Emulsifier Safe, PerkinElmer). Samples were placed on a liquid scintillation counter (Tri-Carb 4910 TR, PerkinElmer) for 5 mins per sample to count radioactivity disintegrations per minute (DPM). Blank samples taken from rats with no radioactivity were also counted with each run to establish background levels. These were always subtracted from the plasma, CSF or tissue sample counts.
Soluene 350
Manufactured by PerkinElmer
Sourced in United States, United Kingdom
Soluene-350 is a liquid scintillation cocktail designed for the dissolution and counting of liquid and solid radioactive samples. It is a clear, colorless solution that provides high efficiency and low background for the measurement of radioactive isotopes in aqueous and organic samples.
Automatically generated - may contain errors
Lab products found in correlation
Hionic fluor, by PerkinElmer (5 mentions)
Ultima gold liquid scintillation cocktail, by PerkinElmer (3 mentions)
Tri carb 4910 tr, by PerkinElmer (3 mentions)
Emulsifier safe, by PerkinElmer (3 mentions)
Acetonitrile, by Thermo Fisher Scientific (3 mentions)
Ethanol, by Merck Group (2 mentions)
Ultima gold, by PerkinElmer (2 mentions)
Soluene350, by Merck Group (2 mentions)
Glacial acetic acid, by Merck Group (2 mentions)
Hionic fluor scintillation cocktail, by PerkinElmer (2 mentions)
Dithiothreitol (dtt), by Fujifilm (2 mentions)
Protease, by Fujifilm (2 mentions)
Tyrosinase, by Merck Group (2 mentions)
Proteinase k, by Fujifilm (2 mentions)
6 m hcl, by Fujifilm (2 mentions)
Benzenesulfonamide, by Merck Group (2 mentions)
Rapid equilibrium dialysis device, by Thermo Fisher Scientific (2 mentions)
3 cc oasis hlb solid phase extraction cartridge, by Waters Corporation (2 mentions)
4 hydroxybenzenesulfonamide, by Merck Group (2 mentions)
Ultima gold scintillation cocktail, by PerkinElmer (2 mentions)
42 protocols using soluene 350
1
Radioactive Tissue and Fluid Analysis
2
Intestinal Glucose Uptake in Diabetic Mice
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Uptake measurements were performed in db/db mice
as described.21 (link) The animals were starved
overnight with free access to water or peptide-containing water. Between
11 and 12 a.m., the mice were killed. The small intestine was removed,
perfused with Krebs–Ringer buffer [25 mM N-(2-hydroxyethyl)piperazine-N′-ethanesulfonic
acid pH 7.4, 108 mM NaCl, 4.8 mM KCl, 1.2 mM KH2PO4, 1.2 mM CaCl2], and stored between 2 and 3 h in
DMEM containing 5 mMd -glucose. For uptake measurements,
the intestine was everted, eight 1 cm long segments of the proximal
jejunum were prepared, and the samples were washed with Krebs–Ringer
buffer. They were incubated for 2 min at 37 °C with Krebs–Ringer
buffer containing 1 mM AMG traced with [14C]AMG in the
absence or presence of 0.2 mM phlorizin. Uptake was stopped and the
segments were washed with ice-cold Krebs–Ringer buffer containing
0.2 mM phlorizin. After solubilizing the samples with tissue solubilizer,
Soluene-350 (PerkinElmer Inc., Waltham, MA) radioactivity was determined
by liquid scintillation counting. In each mouse, eight uptake measurements
were performed, four in the absence and four in the presence of phlorizin,
and mean values of phlorizin inhibited uptake were calculated.
as described.21 (link) The animals were starved
overnight with free access to water or peptide-containing water. Between
11 and 12 a.m., the mice were killed. The small intestine was removed,
perfused with Krebs–Ringer buffer [25 mM N-(2-hydroxyethyl)piperazine-N′-ethanesulfonic
acid pH 7.4, 108 mM NaCl, 4.8 mM KCl, 1.2 mM KH2PO4, 1.2 mM CaCl2], and stored between 2 and 3 h in
DMEM containing 5 mM
the intestine was everted, eight 1 cm long segments of the proximal
jejunum were prepared, and the samples were washed with Krebs–Ringer
buffer. They were incubated for 2 min at 37 °C with Krebs–Ringer
buffer containing 1 mM AMG traced with [14C]AMG in the
absence or presence of 0.2 mM phlorizin. Uptake was stopped and the
segments were washed with ice-cold Krebs–Ringer buffer containing
0.2 mM phlorizin. After solubilizing the samples with tissue solubilizer,
Soluene-350 (PerkinElmer Inc., Waltham, MA) radioactivity was determined
by liquid scintillation counting. In each mouse, eight uptake measurements
were performed, four in the absence and four in the presence of phlorizin,
and mean values of phlorizin inhibited uptake were calculated.
3
Raccoon Dog Hair Melanin Quantification
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Three wild-type (W) and three white-type (B) male raccoon dogs were selected from the experimental base of the Chinese Academy of Agricultural Sciences in Jilin Province. After administering anesthesia, 2 cm × 2 cm of hair coat was cut from the right-hand side of the animals’ backs. The samples were washed, air dried, and sorted, and the guard hair and fluff were placed in valve bags. We improved upon a previous method by transforming the guard and fluff hair into powder, weighing 5 mg of the powder into EP tubes and adding 0.5 mL Soluene®-350 (PerkinElmer) which mix with water at 9:1. We then vortexed and centrifuged the samples for 10 seconds and maintained the temperature at 100 °C for 30 minutes to 1 hour using a thermostat. An appropriate shock was performed to promote hair dissolution. Subsequently, we assessed 200 μL of this solution to determine the absorbance at 500 nm and 650 nm. The solution was weighed according to melanin standards using a precision balance, and Soluene®-350 liquid was added to a stock solution at a concentration of 2 mg/mL. The solution was then diluted to various concentrations, and we assessed 200 μL to determinate the absorbance at 500 nm. A standard curve was then generated to obtain the equation.
4
Hair and Melanin Quantification in Mice
5
Synthesis of POx Block Copolymers
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All materials for the synthesis of POx block copolymers, methanol, ethanol, and sodium lactate were purchased from Sigma-Aldrich (St. Louis, MO). Water and acetonitrile [high-performance liquid chromatography (HPLC) grade] were purchased from Fisher Scientific (Fairlawn, NJ). All materials for the synthesis of POx block copolymers, including methyl trifluoromethanesulfonate (MeOTf), 2-methyl-2-oxazoline (MeOx), 2-ethyl-2-oxazoline (EtOx), 2-n-butyl-2-oxazoline (BuOx), and 2-methoxycarboxyethyl-2-oxazoline (MestOx) were dried by refluxing over calcium hydride (CaH2) under inert nitrogen gas and subsequently distilled before use. Palbociclib-free base, palbociclib-HCl salt form, vismodegib, etoposide, and gemcitabine were purchased from LC Laboratories (Woburn, MA). Sapanisertib was purchased from MedKoo Biosciences (Morrisville, NC). [3H] Palbociclib was purchased from American Radiolabeled Chemicals (St. Louis, MO). Soluene-350 and Ultima Gold LLC scintillation cocktail were purchased from PerkinElmer Life and Analytical Sciences (Waltham, MA).
6
Measuring Cell Proliferation via Radiolabeling
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[3H]Thymidine (1 µCi/ml) was added to TT cells for the last 24 h of siRNA treatment. Radiolabeled cells were extensively washed in ice-cold PBS and dissolved in Soluene-350 (PerkinElmer), after which Ultima Gold LSC Cocktail (PerkinElmer) was added and the radioactivity measured in a PerkinElmer liquid scintillator. The mean±s.e.m. of pooled data from three identical experiments was calculated using a two-sided t-test with Bonferroni correction. A corrected P-value <0.05 was considered significant.
7
Radiolabeled Selegiline Synthesis and Characterization
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Selegiline hydrochloride (Fig. 1
Soluene-350 and Optiphase scintillation cocktails were purchased from Perkin Elmer (PER-FORM Hungária Kft, Budapest, Hungary).
Soluene-350 and Optiphase scintillation cocktails were purchased from Perkin Elmer (PER-FORM Hungária Kft, Budapest, Hungary).
8
Characterization of Radiolabeled Triclocarban
9
Cell Culture Media and Reagents
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Dulbecco’s modified Eagle’s medium (DMEM, 4500 mg/L glucose) was purchased from Sigma-Aldrich (St. Louis, MO, USA); fetal bovine serum (FBS). Penicillin, and streptomycin were purchased from Gibco (Invitrogen, Carlsbad, CA, USA). The tetrazolium salt MTT was obtained from Dojindo Laboratories (Kumamoto, Japan); Soluene-350 was obtained from PerkinElmer, Inc. (Waltham, MA, USA); Blocking One was from Nacalai Tesque (Kyoto, Japan); phosphatase inhibitor was purchased from Roche (Mannheim, Germany); proteinase inhibitor, the Protein Assay kit, and other chemicals were purchased from Wako Pure Chemical Industries, Ltd. Six- and 24-well multiwell plates and 96-well microplates were purchased from Greiner Japan (Tokyo, Japan).
10
Intestinal Glucose Uptake Assay
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Uptake measurements were performed as described [11 (link)]. In brief, rats were starved for 18 h and sacrificed between 10 a.m. and noon. The small intestines were removed and perfused with Krebs-Ringer buffer (25 mM HEPES, 108 mM NaCl, 4.8 mM KCl, 1.2 mM KH2PO4, 1.2 mM CaCl2, pH 7.4, and 37°C). The small intestine was everted and the regions selected for uptake measurements (Figure 1(a) ) were cut into four segments of 1 cm length. The segments were incubated for 2 min at 37°C with Krebs-Ringer buffer containing 10 μM of the SGLT1 specific glucose analogue [14C]α-Methyl-d -glucopyranoside (AMG), with or without 0.2 mM of the SGLT1-specific inhibitor phlorizin. Uptake was stopped by transferring the segments into ice cold Krebs-Ringer buffer containing 0.2 mM phlorizin. After washing with ice cold Krebs-Ringer buffer, the length of the individual segments was measured under a microscope, and the segments were solubilized with Soluene-350 (Perkin Elmer Inc., Waltham, MA, USA). Radioactivity was analyzed by liquid scintillation counting.
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