Dunaliella tertiolecta in batch culture was exposed to Control (f/2 medium) and WAF for a period of seven days in triplicates. WAF was prepared by mixing sterile f/2 growth media and oil (400 μL · L−1). The solution was stirred overnight in the dark and then filtered through a 20 μm nylon mesh to exclude larger oil droplets. The resulting WAF had an oil concentration of 3.25 mg · L−1 measured as estimated oil equivalents according to Wade et al. (2011 ). Briefly, the hydrocarbons in the samples were extracted using dichloromethane, and the fluorescence was measured at 322/376 nm (excitation/emission) wavelength using a Shimadzu spectrofluorometer (RF‐5301PC; Shimadzu, Houston, TX, USA). To understand the oil‐resisting ability of D. tertiolecta, the cells were inoculated at a cell density of 105 cells · mL−1 in control f/2 medium and WAF medium with and without metabolic inhibitors. The metabolic inhibitors used, and their respective target biochemical pathways are listed in Table
Rf 5301pc spectrofluorometer
Manufactured by Shimadzu
Sourced in Japan, United States, Germany, Italy
The RF-5301PC spectrofluorometer is an analytical instrument designed to measure the fluorescence properties of samples. It is capable of quantifying the intensity of fluorescence emitted by a sample when excited by a light source. The core function of the RF-5301PC is to provide accurate and reliable fluorescence data for various applications.
Automatically generated - may contain errors
Lab products found in correlation
Uv 2401 pc spectrophotometer, by Shimadzu (4 mentions)
Uv 1800 spectrophotometer, by Shimadzu (4 mentions)
Uv 2450 spectrophotometer, by Shimadzu (3 mentions)
Spectrometer 4100, by Jasco (2 mentions)
Sonic 6d water bath, by Polsonic (2 mentions)
Ob920 luminescence lifetime spectrometer, by Edinburgh Instruments (2 mentions)
Spectrum two spectrophotometer, by PerkinElmer (2 mentions)
Mcp photomultiplier, by Hamamatsu Photonics (2 mentions)
D8 instrument, by Bruker (2 mentions)
Uv 3600 spectrophotometer, by Shimadzu (2 mentions)
24 well tissue culture plate, by Corning (1 mentions)
J 815 cd spectropolarimeter, by Jasco (1 mentions)
Ft ir perkin elmer spectrometer, by PerkinElmer (1 mentions)
Lambda 25, by PerkinElmer (1 mentions)
Uv transilluminator, by Eppendorf (1 mentions)
Fluorescein 5 maleimide, by Thermo Fisher Scientific (1 mentions)
Tetramethylrhodamine 5 maleimide, by Thermo Fisher Scientific (1 mentions)
Dcfh da, by Merck Group (1 mentions)
Cary 60 uv vis spectrometer, by Agilent Technologies (1 mentions)
C 18 reverse phase high performance liquid chromatography rp hplc, by Beckman Coulter (1 mentions)
126 protocols using rf 5301pc spectrofluorometer
1
Assessing Dunaliella tertiolecta Oil Tolerance
Check if the same lab product or an alternative is used in the 5 most similar protocols
Dunaliella tertiolecta in batch culture was exposed to Control (f/2 medium) and WAF for a period of seven days in triplicates. WAF was prepared by mixing sterile f/2 growth media and oil (400 μL · L−1). The solution was stirred overnight in the dark and then filtered through a 20 μm nylon mesh to exclude larger oil droplets. The resulting WAF had an oil concentration of 3.25 mg · L−1 measured as estimated oil equivalents according to Wade et al. (2011 ). Briefly, the hydrocarbons in the samples were extracted using dichloromethane, and the fluorescence was measured at 322/376 nm (excitation/emission) wavelength using a Shimadzu spectrofluorometer (RF‐5301PC; Shimadzu, Houston, TX, USA). To understand the oil‐resisting ability of D. tertiolecta, the cells were inoculated at a cell density of 105 cells · mL−1 in control f/2 medium and WAF medium with and without metabolic inhibitors. The metabolic inhibitors used, and their respective target biochemical pathways are listed in Table1 .
+ Expand
Dunaliella tertiolecta in batch culture was exposed to Control (f/2 medium) and WAF for a period of seven days in triplicates. WAF was prepared by mixing sterile f/2 growth media and oil (400 μL · L−1). The solution was stirred overnight in the dark and then filtered through a 20 μm nylon mesh to exclude larger oil droplets. The resulting WAF had an oil concentration of 3.25 mg · L−1 measured as estimated oil equivalents according to Wade et al. (2011 ). Briefly, the hydrocarbons in the samples were extracted using dichloromethane, and the fluorescence was measured at 322/376 nm (excitation/emission) wavelength using a Shimadzu spectrofluorometer (RF‐5301PC; Shimadzu, Houston, TX, USA). To understand the oil‐resisting ability of D. tertiolecta, the cells were inoculated at a cell density of 105 cells · mL−1 in control f/2 medium and WAF medium with and without metabolic inhibitors. The metabolic inhibitors used, and their respective target biochemical pathways are listed in Table
2
Fluorescence Studies of bis-ANS Binding
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescence studies of the hydrophobic fluorophore bis-ANS (Sigma, Cat # 65664-81-5) were carried out in a Shimadzu spectrofluorometer (RF-5301PC) as reported (29 (link)). The fluorophore was dissolved in DMSO to a final concentration of 10 mM. The fluorophore was excited at 399 nm, and the emission spectrum was recorded from 420 to 600 nm. To a fixed concentration of N (850 nM) in the reaction buffer (20 mM Tris-HCl pH 8.0, 100 mM NaCl and 3% DMSO), small aliquots of bis-ANS were added from a higher-concentration stock and fluorescence intensity at 485 nm was recorded at each input concentration of bis-ANS. Similarly, N was first incubated with K31 at room temperature for 45 min, followed by the addition of small aliquots of bis-ANS from a higher-concentration stock and fluorescence intensity at 485 nm was recorded. To determine the change in fluorescence signal of bis-ANS due to binding with N, the fluorescence signal of free bis-ANS in the reaction buffer without N was subtracted. The subtracted fluorescence signal was plotted versus each input concentration of bis-ANS.
3
Kinetics of FtsZ Filament Assembly
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The assembly kinetics of FtsZ filaments and bundles with or without ZapA and ZapAL were measured by the light scattering method at room temperature. After FtsZ and the ZapAL or ZapA mixture is quickly mixed with GTP, the changes of the light scattering signal are tracked to obtain their assembly kinetics. Both excitation and emission were set to 340 nm with a Shimadzu RF-5301 PC spectrofluorometer. Each measurement was repeated two or three times.
4
Determining Critical Micelle Concentration of AMPs
5
Fluorescence Characterization of Melanoidins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Example 16
Characterization of Melanoidins by Fluorescence Excitation-emission Spectra
Spectra of different melanoidins (M1, M3, M7 and M9) were collected on a Shimadzu RF-5301PC spectrofluorometer over the excitation range of 210-590 nm and emission range of 220 to 600 nm. Excitation and emission slit of 5 nM, at a high sensitivity, resolution of 2 nm, and increment of 5 nm. Aqueous solutions were 22 mg/L.
The results presented in
6
Sustained Release of GM-CSF from PEG-PLGA Hydrogels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fluorescein
(FITC)-labeled GM-CSF was added to 15, 20, or 25% (w/v) mPEG–PLGA
copolymer aqueous solution to form a hydrogel. The resulting hydrogels
were plated on 24-well tissue culture plates (Corning, Cambridge,
MA) and incubated at 37 °C with PBS. To obtain the release kinetics
of GM-CSF from hydrogel, the releasing media were collected to measure
FITC fluorescence, and fresh PBS was replaced at regular time intervals
for continuous monitoring of protein release. FITC fluorescence (excitation
488 nm, emission 515 nm) was measured by a Shimadzu RF-5301PC spectrofluorometer
(Japan).
(FITC)-labeled GM-CSF was added to 15, 20, or 25% (w/v) mPEG–PLGA
copolymer aqueous solution to form a hydrogel. The resulting hydrogels
were plated on 24-well tissue culture plates (Corning, Cambridge,
MA) and incubated at 37 °C with PBS. To obtain the release kinetics
of GM-CSF from hydrogel, the releasing media were collected to measure
FITC fluorescence, and fresh PBS was replaced at regular time intervals
for continuous monitoring of protein release. FITC fluorescence (excitation
488 nm, emission 515 nm) was measured by a Shimadzu RF-5301PC spectrofluorometer
(Japan).
7
Fluorometric Assay for ALDH Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HsALDH activity was measured using the substrate 6-methoxy-2-naphthaldehyde (5 µM) and the coenzyme NAD+ (100 µM) in 50 mM sodium phosphate buffer (pH 7.5) at 25°C, in the presence of 0.5 mM EDTA and 0.5 mM DTT [26 (link), 27 (link)]. The reaction was initiated by the addition of the enzyme in the reaction mixture at 25°C and monitoring continuously for 5 min. If any fluorescence background drift was there, it was measured before adding the enzyme and was subtracted from the final slope. Fluorescence assays were performed on a Shimadzu RF-5301PC Spectro-fluorometer with excitation and emission wavelengths as 315 nm and 360 nm, respectively. The inner filter effect was nullified by using the following equation (1):
Fcor = Fobs10(Aex+Aem)/2 (1)
Where, Fcor and Fobs are the corrected and observed fluorescence intensity, respectively. Aex and Aem are the absorbance at excitation and emission wavelength, respectively. The reaction velocity was converted in terms of product formation using a standard curve of 6-methoxy-2-naphthoic acid. One unit (U) of enzyme activity was defined in terms of number of micromoles of product produced per min per microgram of the enzyme [28 (link)].
Fcor = Fobs10(Aex+Aem)/2 (1)
Where, Fcor and Fobs are the corrected and observed fluorescence intensity, respectively. Aex and Aem are the absorbance at excitation and emission wavelength, respectively. The reaction velocity was converted in terms of product formation using a standard curve of 6-methoxy-2-naphthoic acid. One unit (U) of enzyme activity was defined in terms of number of micromoles of product produced per min per microgram of the enzyme [28 (link)].
8
Isolation and Characterization of Myelin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Myelin was isolated from whole brain of 3 month-old mice by sucrose density-gradient centrifugation, according to the Norton and Poduslo method (1973 (link)). We used six independent myelin preparations per genotype (n = 3 brains/preparation). Myelin protein content was determined with MicroBCA Protein Assay (Pierce) and monitored by SDS-PAGE and Coomassie blue staining. Myelin (1 mg/ml) was fluorescently labeled by incubation with 12.5 mg/ml of the lipophilic dye 1,1″-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanide perchlorate (DiI; Sigma) for 30 min at 37°C. Excess of DiI was removed by washing with sterile PBS followed by centrifugation (20 min at 24000 g). Myelin fluorescence was determined in a Shimadzu RF-5301PC spectrofluorometer. Excitation wavelength was 545 nm. Fluorescence emission was recorded from 490 to 600 nm. Labeled myelin was stored in small aliquots at −20°C in the dark.
Myelin particle area and total counts were measured using ImageJ software (Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA,http://imagej.nih.gov/ij/ , 1997–2014) and thresholded images of DiI-labeled myelin preparations freshly spread on microscope slides.
Myelin particle area and total counts were measured using ImageJ software (Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA,
9
Intracellular ROS Production Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Intracellular ROS production in extracts of Aloe castellorum and Aloe pseudorubroviolacea treated and untreated HCT-116 cells were estimated through non-fluorescent probe DCFH-DA that can merely pass into the intracellular cell matrix, there it was oxidized into fluorescent dichlorofluorescein (DCF) by the action of produced ROS. Consequently, the fluorescence strength was comparatively proportional to the level of ROS production (Jesudason et al., 2008 (link)). The HCT-116 cells were inoculated (1 × 106 cells/well) into 6-well plate, tested with various concentrations (25, 50 and 75 µg) of extracts Aloe castellorum and Aloe pseudorubroviolacea and placed in CO2 (5%) incubator for 24 h. Then the cells were exposed to 100 µL of DCFH-DA for 10 min at 37 °C. Fluorescence depth was estimated through excitation and emission filters fixed at 485 and 530 nm, respectively (Shimadzu RF-5301 PC spectrofluorometer). The results showed an increased percentage of fluorescence depth.
10
Fluorescence Calibration of Molecular Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A series of dilutions from 1 to 10 µM of the free fluorescent marker and the fluorescent marker in CB[6] in water were prepared to obtain calibration graphs. Fluorescence spectra were measured on a Shimadzu RF-5301 PC spectrofluorometer with excitation and emission wavelengths of 338 nm and 377 nm respectively, through a quartz cuvette with a path length of 1 cm.
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!