Enzymatic activity of DOT1L was assessed by incubating 125 nM of recombinantly expressed and purified GST-DOT1L in the presence of 125 nM (0.28 μCi) 3H-S-adenosyl methionine (NET155250UC; Perkin Elmer), and 0.7 μg of recombinant nucleosomes in HMTase buffer (20 mM Tris pH 7.9, 4 mM EDTA, 1 mM DTT, 0.01% Triton X-100) at a final volume of 26.5 μL. For determination of inhibitor potency, compounds were added to the reaction mixture prior to initiation of the HMTase reaction with 3H-SAM. The HMTase reaction was allowed to proceed for 1 hr at room temperature and was stopped by transferring 5 μL of reaction mixture to P81 filter paper. After drying, filter papers were washed three times with 50 mM NaHCO3 pH 9.0. Filter papers were then dried and radioactivity measured in vials with 10 mL liquid scintillation fluid on a Tri-Carb 2800 TR liquid scintillation counter (Perkin Elmer).
Tri carb 2800tr liquid scintillation counter
Manufactured by PerkinElmer
Sourced in United States
The Tri-Carb 2800TR is a liquid scintillation counter manufactured by PerkinElmer. It is a device used for the detection and quantification of radioactive samples.
Automatically generated - may contain errors
6 protocols using tri carb 2800tr liquid scintillation counter
1
Enzymatic Activity Assay of DOT1L
2
Intracellular Iron Tracing in Plant Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plants were grown for 7 days in plates containing 1/6-strength MS supplemented with 0.8% (w/v) of agar and transferred to plates supplemented with 600 µM FeNaEDTA and 1.67 µCi ml–1 of ferric 55Fe for four days. To avoid the contact between shoots and the media, a glass cover slide was placed between the shoot and the agar surface. After this, the root and shoot tissues were harvested separately into glass vials and washed for 10 min in 5 ml of 1/6-strength MS supplemented with 600 µM Fe(III)-EDTA with gentle shaking. This washing step ensures the removal of 55Fe from the media on the external surface of roots or shoots, while preserving the intracellular 55Fe as well as the 55Fe precipitated into cell walls. The samples were dried for two days at 60 °C before being digested with 0.2 ml perchloric acid at 90 °C for 8 h. The solution was cleared by adding 0.4 ml H2O2 and incubating overnight at 90 °C. For the 55Fe quantification, 4 ml of HionicFluor scintillating mixture (PerkinElmer, Waltham, MA, USA) was added to the samples, and the counts per minute were measured with a Tri-Carb 2800TR liquid scintillation counter (PerkinElmer)78 (link).
3
Measuring Intracellular Drug Accumulation with [3H]-Saquinavir
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Radio-labeled 3H-SQV (specific activity: 1.0 Ci/mM) was obtained from Moravek Biochemicals (Brea, CA, USA) and was used to measure intracellular drug accumulation [7 (link)]. Briefly, HuT78 cells were cultured in 24-well plates (5 × 105 per well) and pre-exposed to CyO2 (0.5 and 1.5 μM) for either 10 min or 30 min, followed by washing off with PBS and addition of 3H-SQV (1.7 pM) and incubation at 37 °C for 2 h. Cells were harvested by centrifugation and extracts obtained by lysing with 1.0 M ammonium hydroxide (NH4OH). Intracellular levels of 3H-SQV were monitored in the lysates, and 100 μL of the lysate was used to measure protein levels by using the BCA protein assay kit (ThermoFisher, Waltham, MA, USA). The remaining 100 μL of the lysate was dissolved in 10 mL of EcoLite scintillation fluid from MP Biomedicals (Santa Ana, CA, USA) and count per minute (CPM) were determined by using a Tri-Carb 2800TR Liquid Scintillation counter (Perkin Elmer, Waltham, MA, USA). In respective samples, data were normalized to the protein contents and presented as CPM/μg of protein.
4
Quantification of Radioactive Tracer in Biological Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The TRA in plasma, whole blood, urine, and feces was determined by liquid scintillation counting. Plasma (0.2 mL) and urine (1 mL) samples were mixed directly with 10 mL liquid scintillation cocktail (Ultima GoldTM, Perkin Elmer Inc., Waltham, MA, USA). To the whole blood samples (0.2 mL), 1 mL Solvable (Perkin Elmer Inc.), 0.1 mL 0.1 M EDTA, and 0.5 mL 30 % hydrogen peroxide were added to dissolve and decolorize the samples. Feces homogenates (0.2 mL) were first dissolved and decolorized using 1 mL Solvable (Perkin Elmer Inc.), 1 mL isopropanol, and 0.4 mL 30 % hydrogen peroxide. The decolorization reaction was started by warming the samples in a shaking water bath of approximately 43 °C, after which the samples were placed in a dark cool place for at least 1 h before liquid scintillation cocktail (10 mL) was added. Samples were counted on a Tri-Carb® 2800TR liquid scintillation counter (Perkin Elmer Inc.). Quench correction was applied with a calibration curve of quenched radioactive reference standards. Samples were counted to a sigma 2 counting error of 5 % or for 60 min at most. The lower limit of quantitation (LLOQ) was approximated to be 1.2 to 1.4 ng-eqv/g using the counting error as described previously [18 (link), 19 (link)].
5
Primary Production Measurement by 14C Incubation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary production (PP) was conducted according to Parsons et al.61 by 14C incubation method61 . Seawater samples were collected from surface (Table 1 ) or five depths of the euphotic layer (Table 2 ), then prescreened through 200 μm mesh and filled into 250 mL acid-cleaned polycarbonate carboy (Nalgene; USA). Each sample was inoculated with 10 μCi NaH14CO3 before incubation, and carboys which used for incubate subsurface samples were covered with the neutral density filter to simulate the in situ irradiance. Carboys were incubated in an on-deck incubator cooled by running water pumped up from a 5 m depth. After 4 h incubation, seawater samples were filtered through 25 mm diameter GF/F filters, and then the filters were placed in the aluminum foil bags and stored in the −20 °C freezer until analysis. In the laboratory, filters were fumed with HCl to remove residual inorganic carbon before they were moved into the scintillation vials, next the 10 mL scintillation cocktail (Ultima Gold; PerkinElmer; USA) were added. The activity of radioactive was counted using a Tri-Carb 2800TR liquid scintillation counter (Perkin-Elmer; USA).
6
Quantitative Radiotracer Measurement
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The TRA in plasma, whole blood, urine and feces was determined by liquid scintillation counting (LSC). Plasma (0.2 mL) and urine (1 mL) samples were directly mixed with 10 mL liquid scintillation cocktail (Ultima Gold™ . , Perkin Elmer Inc., Waltham, MA, USA). To the whole blood samples (0.2 mL) 1 mL Solvable (Perkin Elmer Inc.), 0.1 mL 0.1 M EDTA, and 0.5 mL 30% hydrogen peroxide was added to dissolve and to decolorize the samples. Feces homogenates (0.2 mL) were first dissolved and decolorized using 1 mL Solvable (Perkin Elmer Inc.), 1 mL isopropanol, and 0.4 mL 30% hydrogen peroxide. The decolorization reaction was started by warming the samples in a shaking water bath of approximately 43 °C after which the samples were placed at a dark cool place for at least 1 h before adding liquid scintillation cocktail. Samples were counted on a Tri-Carb® 2800TR Liquid Scintillation Counter (Perkin Elmer Inc.). Quench correction was applied with a calibration curve of quenched radioactive reference standards. Samples were counted to a sigma 2 counting error of 1% or for a maximum of 60 min.
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!