Whole placentas and minced fetuses were digested in Biosol (National Diagnostics, Hull, UK) for ≥ 1 week at 55°C. Beta emissions of maternal plasma and of fetal and placental digestates were measured using a 300 SL Liquid Scintillation Counter (LabLogic, Sheffield, UK). Radioactivity in the fetuses, placentas and maternal plasma was used to calculate either placental clearance of MeGlu and MeAIB (μl min–1 g–1 of placenta) or tracer accumulation expressed (g–1 of fetus or placenta), as described previously (Sibley et al. 2004 ).
Biosol
Manufactured by National Diagnostics
Sourced in United Kingdom
Biosol is a laboratory instrument designed for the analysis and quantification of biological samples. It utilizes a spectroscopic detection method to measure the absorbance or fluorescence of samples, enabling the determination of various analytes in a range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Bioscint, by National Diagnostics (3 mentions)
Tri carb 2910tr liquid scintillation analyzer, by PerkinElmer (1 mentions)
Ab23722, by Abcam (1 mentions)
Ab125145, by Abcam (1 mentions)
Ab192603, by Abcam (1 mentions)
Ab150687, by Abcam (1 mentions)
Tnfα elisa, by Thermo Fisher Scientific (1 mentions)
Thp 1 cell, by American Type Culture Collection (1 mentions)
Glutaraldehyde, by Polysciences (1 mentions)
Ecoscint, by National Diagnostics (1 mentions)
5 fluorouracil, by Merck Group (1 mentions)
Nile red, by MP Biomedicals (1 mentions)
Sulforhodamine b, by Merck Group (1 mentions)
14c 5 fu, by Moravek Biochemicals (1 mentions)
Hepes, by Merck Group (1 mentions)
Sodium borohydride, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
Sodium azide, by Merck Group (1 mentions)
Glyoxal, by Merck Group (1 mentions)
9 protocols using biosol
1
Placental Tracer Clearance Measurement
2
Measuring Materno-Fetal Tracer Transfer
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Unidirectional materno-fetal transfer of non-metabolizable radioactive tracers was measured as described by Coan et al.27 (link). Briefly, pregnant mice were anesthetized with an intraperitoneal injection (0.4 ml) of fentanyl/fluanisone and midazolam solutions in water (1:1:2 water, Janseen Animal Health). After the maternal jugular vein was exposed, a 100 μl bolus of PBS containing 3.5 μCi (1 Ci = 37GBq) of [14C]methyl amino-isobutyrate (MeAIB) (specific activity 50.5 mCi/mmol) or 3.5 μCi of [14C]-methyl-D-glucose (specific activity 56.4 mCi/mmol) was injected into the jugular vein via a short length of tubing attached to a 27 gauge needle and connected to a 1 ml syringe. At times up to 4 min after tracer injection, the mice were killed and their conceptuses were dissected via hysterectomy. The fetuses were lysed overnight in 2 ml of Biosol (National Diagnostics) at 55 °C. Fetal sample fractions were then added to the appropriate tubes for β counting (Packard Tri-Carb 1900). The radioactive counts of each fetus were used to calculate the amount of radioisotope transferred per gram of placenta or fetus.
3
In Vivo Fatty Acid Uptake in Clenbuterol-Treated Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In vivo fatty acid uptake was examined in WT male mice treated with clenbuterol (30 mg/l) or regular drinking water for 5 days. Mice were fasted overnight (15 h) and injected i.p. with 200 μl of olive oil containing 2 μCi of (R)-2-bromopalmitic acid [9,10-3H] ([3H]-BROMO), a partially metabolizable long-chain fatty acid tracer (American Radiolabeled Chemicals, St. Louis, MO). Tissues (~150 mg) were collected 1 h later and solubilized in 1 ml of Biosol (National Diagnostics, Charlotte, NC) for 3 h in a 50 °C water bath, followed by another 1 h incubation in the presence of 0.3 ml 30% hydrogen peroxide. Samples were mixed with 10 ml of scintillation solution (Bioscint, National Diagnostics, Charlotte, NC) and counted using a Tri-Carb 2910 TR liquid scintillation analyzer (Perkin Elmer, Waltham, MA). Data were normalized to mg of tissue.
4
Functionalized POZ Polymers for Biomedical Applications
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mono- and diamino-functionalized POZ, (10 kDa) were chosen as optimized polymers; these compounds were purchased from Ultroxa. 5(6)-carboxyfluorescein N-hydroxysuccinimide ester (≥80%) was purchased from Sigma. Antibodies were purchased from Abcam: anti-BSA antibody (ab192603), anti-AGE antibody (ab23722), anti-CML antibody (ab125145). Glutaraldehyde was purchased from Polysciences. Biosol was purchased from National Diagnostics. Pharmaceutical grade human serum albumin (HSA), used in the clinical grade trileaflet BHV experiments, was purchased from Octapharma. THP-1 cells, a monocyte/macrophage cell line, were purchased from ATCC. RPMI medium was purchased from Cell Culture Technologies. TNF-α ELISA was obtained from Invitrogen. Cellulose dialysis membrane was purchased from Spectrum Labs (10768-700). The Von Kossa staining kit was obtained from Abcam (ab150687). All chemicals were purchased from Sigma Aldrich, unless otherwise stated.
5
Glucose Uptake Kinetics in Germinating Conidia
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
[1,2-3H]2-deoxy-D-glucose (MP Biomedicals, Irvine CA)), at 1 μCi/ml, was added to 15 ml of 5 hr germinating conidia upon their transfer to DS conditions. Aliquots were removed after 15, 45, and 75 min of continued incubation. The pelleted cells from 150 μl of culture (in triplicate) were solubilized in BioSol (National Diagnostics, Atlanta, GA), according to the manufacturer’s instructions, and were mixed with scintillation fluid (Bio-Safe II, RPI, Mount Prospect, IL) for counting. The top 150 μl supernatant from 500 μl of pelleted culture were directly mixed with scintillation fluid and counted. The percentage uptake was determined by dividing the average pellet-associated tritium counts by the sum of the pellet-associated and supernatant counts, which together approximated 0.15 μCi.
6
Radioactive Uptake Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
5-Fluorouracil (5-FU), estradiol (EST) and sulforhodamine B (SRB) were purchased from Sigma-Aldrich, St. Louis, MO. Nile Red (NR) was purchased from MP Biomedicals, Solon, OH. 14C-5-FU and 3H-EST were purchased from Moravek Biochemicals and Radiochemicals, Brea, CA. Biosol (tissue solubilizer), Bioscint and Ecoscint (scintillation cocktails) were purchased from National Diagnostics, Atlanta, Georgia. Isoflurane was purchased from VetOne, Boise, ID. All other reagents were purchased from Sigma-Aldrich, St. Louis, MO, USA.
7
Bovine serum albumin-based radiolabeled glucose assay
8
Maternal-Fetal Tracer Transfer Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The unidirectional materno-fetal clearance of the non-metabolisable radioactive tracers, 14C-methyl amino-isobutyric acid (MeAIB) and 3H-methyl D-Glucose (MeG) were measured under anesthesia with fentanyl-fluanisone (hypnorm):midazolam (hypnovel) in sterile water (1:1:2, Jansen Animal Health) (Sibley et al., 2004 (link)). A 200 μl bolus containing 3.5 µCi, of MeAIB (NEN NEC-671; specific activity 1.86GBq/mmol, Perkin Elmer, USA) and 3.5 µCi MeG (NEN NEC-377; specific activity 2.1GBq/mmol) in physiological saline (0.9% wt/vol) was injected into the maternal jugular vein. Two minutes after tracer injection, the dam was killed by cervical dislocation, uteri were collected and fetal and placental weights recorded. Entire litters of placentas were collected for morphological analyses or snap frozen in liquid nitrogen for quantification of gene expression. Fetuses were decapitated, entire fetal tails taken for DNA genotyping and then fetuses minced and lysed at 55°C in Biosol (National Diagnostics, Atlanta, USA). Fetal lysates were measured for beta emissions by liquid scintillation counting (Optiphase Hisafe II and Packard Tri-Carb, 1900; Perkin-Elmer USA) and radioactivity (DPM; disintegrations per minute) in the fetuses was used to calculate transfer relative to the estimated placental surface area or to fetal weight.
9
Placental Transport and Gene Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pregnant F1 females, and untreated females mated with F1 males were anaesthetized on D19 of pregnancy and unidirectional materno-fetal clearance of 14C-methylaminoisobutyric acid (100µl, NEN NEC-671, specific activity 1.86 GBq mmol -1 ) determined as described previously (Vaughan et al. 2012) . A cardiac blood sample was taken up to 4 minutes after injection of the tracer and the mother was killed by cervical dislocation. Plasma was separated by centrifugation for liquid scintillation counting and corticosterone measurement.
The gravid uterus was removed and individual fetuses and placentae dissected and weighed.
Fetuses were lysed (5ml Biosol, National Diagnostics, UK at 55°C) for liquid scintillation counting. The placenta closest in weight to the mean of each litter was halved and fixed in formaldehyde (4% in 0.1M HEPES) for histological processing and stereological determination of volumes of the Lz and Jz, responsible for the transport and endocrine functions of the placenta (Vaughan et al. 2012) . The placenta second closest to the mean was snap frozen in liquid nitrogen for later gene expression analysis. Expression of System A amino acid transporter isoforms, Slc38a1, Slc38a2 and Slc38a4, was determined relative to Hprt1 and Gapdh using quantitative RT-PCR and the ΔΔCt method (Vaughan et al. 2012) .
The gravid uterus was removed and individual fetuses and placentae dissected and weighed.
Fetuses were lysed (5ml Biosol, National Diagnostics, UK at 55°C) for liquid scintillation counting. The placenta closest in weight to the mean of each litter was halved and fixed in formaldehyde (4% in 0.1M HEPES) for histological processing and stereological determination of volumes of the Lz and Jz, responsible for the transport and endocrine functions of the placenta (Vaughan et al. 2012) . The placenta second closest to the mean was snap frozen in liquid nitrogen for later gene expression analysis. Expression of System A amino acid transporter isoforms, Slc38a1, Slc38a2 and Slc38a4, was determined relative to Hprt1 and Gapdh using quantitative RT-PCR and the ΔΔCt method (Vaughan et al. 2012) .
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!