Liquid scintillation analyzer

Manufactured by PerkinElmer

Sourced in United States, Norway

The Liquid Scintillation Analyzer is a laboratory instrument designed to detect and quantify radioactive samples. It measures the light emitted by a liquid scintillation cocktail when it interacts with radioactive particles. This device is used to analyze the radioactive content of various samples in a wide range of scientific applications.

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Lab products found in correlation

3h thymidine, by PerkinElmer (3 mentions) Liquid scintillation cocktail, by Thermo Fisher Scientific (2 mentions) Tri carb 2910 tr, by PerkinElmer (2 mentions) Deae filtermat, by PerkinElmer (2 mentions) S adenosyl l methyl 3h methionine sam, by PerkinElmer (2 mentions) Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions) Mn gf 5, by Macherey-Nagel (1 mentions) Pd 10 desalting column, by GE Healthcare (1 mentions) Cytochalasin b, by Merck Group (1 mentions) Tnt t7 quick coupled transcription translation system, by Promega (1 mentions) Proteinase, by Roche (1 mentions) And phosphatase inhibitor, by Roche (1 mentions) Gamma 32p atp, by PerkinElmer (1 mentions) Cellulose filter, by Cytiva (1 mentions) Trypsin edta, by Thermo Fisher Scientific (1 mentions) Trichloroacetic acid, by Merck Group (1 mentions) Proteinase inhibitor cocktail, by Roche (1 mentions) 1 2 3h cholesterol, by PerkinElmer (1 mentions) Lc ms, by Agilent Technologies (1 mentions) Scintillation fluid, by PerkinElmer (1 mentions)

27 protocols using liquid scintillation analyzer

Pyruvate Uptake Assay in Salmonella Typhimurium

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To determine the uptake of pyruvate by S. Typhimurium, a transport assay was performed with radiolabeled pyruvate. Cells were grown under agitation at 37 °C in LB and harvested in mid-log phase. Cells were pelleted at 4 °C, washed twice, and resuspended in transport buffer (1 g/L (NH₄)₂SO₄, 10 g/L K₂HPO₄, 4.5 g/l KH₂PO₄, 0.1 g/L MgSO₄, pH 6.8) to an absorbance of 5 at 420 nm, equivalent to a total protein concentration of 0.35 mg/mL. Uptake of ¹⁴C-pyruvate (55 mCi/mmol, Biotrend, Köln, Germany) was measured at a total substrate concentration of 10 µM at 18 °C. At various time intervals, transport was terminated by the addition of ice-cold stop buffer (100 mM potassium phosphate, pH 6.0, 100 mM LiCl) followed by rapid filtration through membrane filters (MN gf-5, 0.4 μm nitrocellulose, Macherey Nagel, Düren, Germany). The filters were dissolved in 5 mL scintillation fluid (MP Biomedicals, Eschwege, Germany), and radioactivity was determined in a liquid scintillation analyzer (PerkinElmer, Waltham, MA, USA).

Paulini S., Fabiani F.D., Weiss A.S., Moldoveanu A.L., Helaine S., Stecher B, & Jung K. (2022). The Biological Significance of Pyruvate Sensing and Uptake in Salmonella enterica Serovar Typhimurium. Microorganisms, 10(9), 1751.

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Measuring DNA-Protein Crosslinks by Alkaline Elution

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Alkaline elution was used to assess cellular TOP1cc by measuring DPC as described (4 (link),31 (link)-33 (link)). Before alkaline elution and drug treatments, genomic DNA of CCRF-CEM cells was radiolabeled with 0.02 μCi/ml [³H]-thymidine for one to two doubling times at 37°C and chased in non-radioactive medium overnight. Cells were treated with the indicated concentrations of LMP compounds or CPT for 1 h before scraping and alkaline elution. For reversal experiments, the cells were cultured in drug-free medium for the indicated times. Radioactivity in all fractions was measured with a liquid scintillation analyzer (PerkinElmer Life and Analytical Sciences), and DPC frequency, which reflects TOP1cc was determined as published (4 (link),31 (link)-33 (link)).

Marzi L., Agama K., Murai J., Difilippantonio S., James A., Peer C.J., Figg W.D., Beck D., Elsayed M.S., Cushman M, & Pommier Y. (2018). Novel fluoroindenoisoquinoline non-camptothecin topoisomerase I inhibitors. Molecular cancer therapeutics, 17(8), 1694-1704.

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Methyltransferase Assay for DNA Methylation

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The methyltransferase assay was carried out at 30°C for one hour in a total volume of 25 μl containing 1.5 μl of S-adenosyl-l-[methyl-³H] methionine (SAM) (14.4 Ci/mmol; PerkinElmer), 1.5 μl substrate DNA (12 repeats of TAC, annealed to form dsDNA, 15 μM), and 0.2 μM AtDRM2 full length (59–626) or DRM2 methyltransferase (DRM2^CAT, 269–626) proteins, 1 μM His-tag UVR8 or GFP proteins in assay buffer (20 mM MOPS [pH 7.0], 1 mM DTT, 5 mM EDTA, 200 μg/ml BSA, and 5% glycerol). The reactions were stopped by adding 1 μl of cold SAM (NEB). A total of 11 μl from each reaction was applied onto DEAE Filtermat (PerkinElmer,1450–522) and washed two times with 200 mM ammonium bicarbonate, two times with water, and two times with ethanol. The paper was dried and placed into 4 mL of liquid scintillation cocktail (Fisher Scientific) and the activity was measured by Liquid Scintillation Analyzer (PerkinElmer, Tri-Carb 2910 TR).

Jiang J., Liu J., Sanders D., Qian S., Ren W., Song J., Liu F, & Zhong X. (2021). UVR8 interacts with de novo DNA methyltransferase and suppresses DNA methylation in Arabidopsis. Nature plants, 7(2), 184-197.

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Glucose Uptake in Adipocytes

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Adipocytes were incubated in DMEM containing 0.2% BSA for 4 h and washed two times with Krebs-Ringer Hepes buffer (136 mM NaCl, 4.7 mM KCl, 1.25 mM CaCl₂, 1.25 mM MgSO₄, 20 mM Hepes, pH 7.4). The cells were incubated in KRH buffer with or without 10 ng/mL insulin at 37 °C for 15 min. Glucose uptake reaction was initiated by addition of 0.5 μCi/mL 2-deoxy-d-[1-³H(N)] glucose as the final concentration in KRH buffer. After 10 min, the cells were quickly washed two times with ice-cold KRH buffer to terminate the reaction. The cells were lysed with 0.5 N NaOH and the radioactivity was determined using a Liquid Scintillation Analyzer (PerkinElmer, Inc., Waltham, MA, USA).

Lee A., Choi K.M., Jung W.B., Jeong H., Kim G.Y., Lee J.H., Lee M.K., Hong J.T., Roh Y.S., Sung S.H, & Yoo H.S. (2017). Enhancement of Glucose Uptake by Meso-Dihydroguaiaretic Acid through GLUT4 Up-Regulation in 3T3-L1 Adipocytes. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 22(9), 1423.

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Radioactive Retinol Uptake Assay

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Radioactive 11,12-³H(N)-retinol was purchased from PerkinElmer. For the in vitro retinol uptake assay, SAA1 was incubated with ³H-retinol at a 3:1 molar ratio at 4°C for at least 2 hours in serum-free media containing protease inhibitors (Roche). The SAA1-retinol complex was purified using PD-10 desalting column (GE Healthcare). Cells were incubated with 480 nM of SAA1-³H-retinol complex or free ³H-retinol in the serum-free media at 37°C and 5% CO₂ (Fig. 2F). Cells were incubated with 600 nM SAA1–³H-retinol complex with and without RAP in serum-free media at 4°C for 1 hour, washed twice with cold media, and incubated at 37°C and 5% CO₂ for 3 hours (Fig. 2G) and for 2 hours (Fig. 4C). The reactions were stopped by washing the cells with ice-cold DPBS and solubilizing the cells in Biosol solution (National Diagnostics). For the in vivo retinol uptake assay, ³H-retinol was diluted in corn oil and fed to mice (1 μCi/mouse). Fifteen to eighteen hours later, immune cells were isolated from the small intestine and CD11c⁺ cells were isolated by magnetic sorting. Cells (2×10⁵) were solubilized in 0.5 ml of Biosol solution. A 3.5-ml volume of Biosinct solution (National Diagnostics) was mixed with each sample and radioactivity was measured with a liquid scintillation analyzer (PerkinElmer) and normalized to cell number.

Bang Y.J., Hu Z., Li Y., Gattu S., Ruhn K.A., Raj P., Herz J, & Hooper L.V. (2021). Serum amyloid A delivers retinol to intestinal myeloid cells to promote adaptive immunity. Science (New York, N.Y.), 373(6561), eabf9232.

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Glucose Uptake Assay in Yeast

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Candida albicans and S. cerevisiae were cultured as described in “Yeast strains and AgNPs treatment
conditions” and diluted with fresh media containing half the amount of dextrose of the original Sabouraud dextrose broth and YPD broth, respectively, to an OD₆₀₀ of 0.2. Candida albicans and S. cerevisiae were cultured for 1 hour at 37°C and at 30°C, respectively, in the presence or absence of 5 mM NAC. Then, 5 nm AgNPs and tritium-labeled 2-deoxyglucose (2-DG) (final 2.5 μCi/mL; Perkin Elmer, Boston, MA, USA) were added to the culture. After 30 minutes, the cells in 1 mL of culture were harvested by centrifugation and washed three times with cold PBS containing the glucose transporter inhibitor cytochalasin B (2.5 μg/mL; Sigma-Aldrich) by centrifugation at 11,400×g, at 4°C for 5 minutes. The cells were then lysed in 400 μL RIPA buffer containing 1% SDS and mixed with 8 mL of scintillation solution. The radioactive count was determined using a liquid scintillation analyzer (Perkin Elmer).

Lee B., Lee M.J., Yun S.J., Kim K., Choi I.H, & Park S. (2019). Silver nanoparticles induce reactive oxygen species-mediated cell cycle delay and synergistic cytotoxicity with 3-bromopyruvate in Candida albicans, but not in Saccharomyces cerevisiae. International Journal of Nanomedicine, 14, 4801-4816.

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Flotillin Lipid Binding Assay

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PCR products with T7 primer and kozac sequences of GFP, flotillin1-GFP, flotillin2-GFP, and different domains of flotillins with C-terminal GFP-tags were used for in vitro transcription/translation (T7 TNT Quick Coupled Transcription/Translation Systems, Promega). The different truncation mutants shown in Fig 4 correspond to residues 3–185 in Flot 1 SPFH, 186–427 Flot 1 flot, 84–266 Flot 1 PHB, 267–467 Flot1 C, 5–188 in Flot 2 SPFH, 189–427 Flot 2 flot, 87–268 Flot 2 PHB, 269–467 Flot 2 C. Protein mixture was diluted with 3% bovine serum albumin (BSA) in PBS, and 0.5 μCi/ml ³H-Sph and magnetic anti-GFP microbeads (Miltenyi Biotec) were added for 1 h incubation on ice. The beads were bound on magnetic columns, washed with BSA/PBS and eluted by removing the magnet. Bound ³H-Sph was analysed by Liquid Scintillation Analyzer (Perkin Elmer). For lipid competition assay, 10 μM of Sph, S1P, or Cer was added to the translated proteins 5 min before adding ³H-Sph and anti-GFP microbeads.

Riento K., Zhang Q., Clark J., Begum F., Stephens E., Wakelam M.J, & Nichols B.J. (2018). Flotillin proteins recruit sphingosine to membranes and maintain cellular sphingosine-1-phosphate levels. PLoS ONE, 13(5), e0197401.

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Cell Proliferation Assay with Tritiated Thymidine

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The proliferation of the tested cells was estimated as [³H]-thymidine incorporation into the DNA of the cells treated with the studied samples [40 (link)]. Firstly, cells (1 × 10⁴ per well) were placed in 24-well plates and were allowed to adhere for 24 h. Afterwards, the cells were treated with 1 mL of growth medium containing 0.5 μCi [³H]-thymidine and various concentrations of each sample (HP1 and HP6-HP7; compounds 1 and 7; and ZAP). cPT was used as a positive control. After incubation for 24 h and 48 h at 37 °C, the cells were rinsed with PBS and solubilized with 1 mL of 0.1 M NaOH (sodium hydroxide) containing 1% SDS (sodium dodecyl sulfate). Then, radioactivity was recorded in a Liquid Scintillation Analyzer (Perkin-Elmer, Waltham, MA, USA) after correction to 9 mL with scintillation liquid.

Strawa J.W., Jakimiuk K., Szoka Ł., Brzezinski K., Drozdzal P., Pałka J.A, & Tomczyk M. (2022). New Polymethoxyflavones from Hottonia palustris Evoke DNA Biosynthesis-Inhibitory Activity in An Oral Squamous Carcinoma (SCC-25) Cell Line. Molecules, 27(14), 4415.

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AMPK Activity Assay in HUVEC Cells

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HUVEC stably expressing EV or FLAG-K1 were incubated in EBM-2 without serum and growth factors for 24 hours. Either 5 uM compound C or DMSO (0.05%) control was added at the start of starvation. Cells were washed with cold PBS and then lysed with a buffer containing 50mM Tris-HCL pH 7.4, 1 mM EDTA, 1 mM EGTA, 250 mM mannitol, 1% Triton X-100 and proteinase (Roche) and phosphatase inhibitors (Roche). Lysates were then clarified by centrifugation. For the AMPK activity assay, reagents were purchased from SignalChem and the manufacturer’s protocol followed. Briefly, 10 μLs of cell lysate was incubated with 5 μL of 1 mg/mL SAMS or peptide substrate solution, 5 μLs 0.5 mM AMP solution and 5 μLs γ-32P-ATP assay cocktail. Gamma-32P-ATP was purchased from Perkin Elmer. The mixture was incubated at room temperature for 30 minutes and then 20 μLs was added to phosphocellulose paper and washed 2 times in 1% phosphoric acid solution. Counts per minute (cpm) were acquired using a PerkinElmer liquid scintillation analyzer.

Anders P.M., Zhang Z., Bhende P.M., Giffin L, & Damania B. (2016). The KSHV K1 Protein Modulates AMPK Function to Enhance Cell Survival. PLoS Pathogens, 12(11), e1005985.

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Renal Cortical Slice Estrone Sulfate Uptake

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The decapsulated kidneys were placed into freshly-oxygenated ice-cold modified Cross and Taggart saline buffer (contain: 95 mM NaCl, 80 mM mannitol, 5 mM KCl, 0.74 mM CaCl₂, and 9.5 mM Na₂HPO₄, pH 7.4). Thin renal cortical slices (≤0.5 mm; 5–15 mg, wet weight) were cut using a Stadie-Riggs microtome and incubated in 1 ml of buffer containing 50 nM [³H] estrone sulfate (ES), a prototypical organic anion that is preferentially transported by Oat3^{35 (link),36 (link)}, to enable an uptake study for 30 mins at room temperature. At the end of the uptake period, the slices were washed in 0.1 M MgCl₂, blotted on filter paper, weighed and dissolved in 0.4 ml of 1 M NaOH, and neutralized with 0.6 ml of 1 N HCl. Five renal cortical slices were used for each rat (5–6 rats per group). The radioactivity was measured using a liquid scintillation analyzer (Perkin Elmer, MA, USA). The transport of ES was calculated as tissue to medium (T/M) ratio.

T / M ratio = dpm / g tissue \div dpm / ml medium

Thongnak L., Pongchaidecha A., Jaikumkao K., Chatsudthipong V., Chattipakorn N, & Lungkaphin A. (2017). The additive effects of atorvastatin and insulin on renal function and renal organic anion transporter 3 function in diabetic rats. Scientific Reports, 7, 13532.

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