Turbidity assays were performed on a UV-Vis spectrophotometer (Cary 100 Bio; Varian) equipped with a multicell Peltier temperature controller. Samples (in quartz cuvettes with 1 cm path length; Thorlabs) were first equilibrated above the transition temperature, and the instrument was blanked. Samples were then cooled at a rate of 1 °C/min until reaching 4 °C. Throughout the experiment, absorbance was measured at λ = 600 nm every 0.5 °C. The samples changed from clear solutions to turbid suspensions upon cooling below the UCST.
Quartz cuvette
Manufactured by Thorlabs
Quartz cuvettes are transparent containers designed for spectroscopic analysis. They are made of high-quality quartz material, which allows for the transmission of light across a wide range of the electromagnetic spectrum, including ultraviolet and visible wavelengths. Quartz cuvettes are commonly used in various analytical techniques, such as absorption, fluorescence, and UV-Vis spectroscopy, to hold samples during measurement.
Automatically generated - may contain errors
Lab products found in correlation
Cary 100 bio, by Agilent Technologies (1 mentions)
Dh 2000 bal deuterium halogen light source, by OceanOptics (1 mentions)
Hr2000 spectrometer, by OceanOptics (1 mentions)
Aunps, by Merck Group (1 mentions)
Acetone, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Isopropyl alcohol ipa, by Merck Group (1 mentions)
Synergy h1, by Agilent Technologies (1 mentions)
Agilent 8453 uv visible spectrophotometer, by Agilent Technologies (1 mentions)
Cary 6000 spectrometer, by Agilent Technologies (1 mentions)
Cary 6000i, by Agilent Technologies (1 mentions)
6 protocols using quartz cuvette
1
Turbidity Assay of UCST Transitions
2
Characterization of Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The AuNPs were purchased from Sigma Aldrich (St. Louis, MO, USA) in 25 mL bottles in 0.1 mM PBS. The studied particle diameters were 20 nm (product no 753610), 40 nm (product no 753637), 60 nm (product no 753653), 80 nm (product no 753661), and 100 nm (product no 753688). Methanol (34885-1L), Acetone (product number 179124), Isopropyl alcohol (IPA) (product number W292907), and Acetonitrile (product number 271004) were also purchased from Sigma Aldrich. Quartz cuvettes (product name CV10Q700F) for UV-VIS absorbance measurements were purchased from Thorlabs. The experimental UV-VIS data were collected using an Ocean Insight (Geograaf, EW Duiven, The Netherlands) HR2000+ spectrometer (product name HR2000CG, Ocean Insight). An Ocean Insight CUV-ALL-UV cuvette holder (product name CUV-UV, Ocean Insight) was used. The light source used was a DH-2000 BAL deuterium halogen light source (product number DH-2000-BAL, Ocean Insight).
3
Comparison of HMU and Modified CHMU Spectra
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The absorbance spectra of the HMU standard and the modified carboxy-HMU (CHMU) were compared to determine similarities between the two fluorophores. The HMU standard and the CHMU were both dissolved in stop solution to a final concentration of 0.5 μmol/l (Ɛ = 13,300 M-1cm-1 at 390 nm) and measured using an Agilent 8453 UV–visible spectrophotometer (Agilent Technologies, Santa Clara, CA). Seven hundred microliters of each solution were added to quartz cuvettes (Thorlabs Inc., Newton, NJ) and the absorbance characteristics were analyzed at wavelengths ranging from 300 – 600 nm that included the expected absorbance maxima at 390 nm, in increments of 1 nm. The experiment was repeated three times with measurements taken in triplicate for each solution. The fluorescence emission spectra for 0.5 μmol/l of HMU standard and CHMU were also analyzed using the BioTek Synergy H1 monochromator-based multi-mode plate reader (Biotek, Winooski, VT). Seventy microliters of each solution were added in triplicate to individual wells of a black half-area 96-well microtiter plate. Fluorescence was measured using an excitation wavelength set to 390 (±9) nm and emission spectra obtained at wavelengths ranging from 420 – 600 nm.
4
Optical Spectroscopic Characterization of Fluorescent Probes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples (1500 μL)
were used in a quartz cuvette (Thorlabs, Newton, NJ) with an optical
path (l) of 1 cm. The temperature of the cell compartment
was set at 25 °C, by a built-in Peltier cooler (Varian). Absorption
data were recorded at 25 °C using 1 nm band pass, 1 nm resolution,
and 200 nm/min scanning speed. Fluorescence intensity measurements
were carried out at the absorption maximum of each compound, employing
an excitation and emission band pass of 2.5 nm, 120 nm/min scan rate
and 600 V PMT detector voltage. For every sample, the absorbance at
the wavelength maximum was recorded and subsequently employed to normalize
the corresponding fluorescence emission intensity.
Fluorescence
emission calibration curves of the three probes were obtained in toluene–PEG
methyl ether methacrylate mixtures at a final concentration of 400
nM.
The time-transient fluorescence emission measurements were
obtained
by adding the appropriate volume of a stock solution of the fluorophores
(C1 , Glu-C1, and Oct-C1 ) in
hotR8 toluene solution (1% w/w) for a final concentration
of 200 nM. The cuvette was kept sealed with a Teflon cap throughout
the duration of the experiment to ensure no variations in concentration.
were used in a quartz cuvette (Thorlabs, Newton, NJ) with an optical
path (l) of 1 cm. The temperature of the cell compartment
was set at 25 °C, by a built-in Peltier cooler (Varian). Absorption
data were recorded at 25 °C using 1 nm band pass, 1 nm resolution,
and 200 nm/min scanning speed. Fluorescence intensity measurements
were carried out at the absorption maximum of each compound, employing
an excitation and emission band pass of 2.5 nm, 120 nm/min scan rate
and 600 V PMT detector voltage. For every sample, the absorbance at
the wavelength maximum was recorded and subsequently employed to normalize
the corresponding fluorescence emission intensity.
Fluorescence
emission calibration curves of the three probes were obtained in toluene–PEG
methyl ether methacrylate mixtures at a final concentration of 400
nM.
The time-transient fluorescence emission measurements were
obtained
by adding the appropriate volume of a stock solution of the fluorophores
(
hot
of 200 nM. The cuvette was kept sealed with a Teflon cap throughout
the duration of the experiment to ensure no variations in concentration.
5
UV-Vis Absorbance Measurement of GtACR1
6
Protein Absorption Measurements in Lipid Membranes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protein samples for absorption measurements were prepared as previously reported 18 , 19 , 20 using approximately 100 µg of the protein in the form of reconstituted ECPL lipid membranes as described above. The samples were washed followed by centrifugation at least three times in approximately 0.1 mL of buffer (pH 3 buffer: 5 mM NaH2PO4, 10 mM NaCl, 10 mM MES; pH 5 buffer: 5 mM NaH2PO4, 10 mM NaCl, 10 mM MES; pH 7.3 buffer: 50 mM NaCl, 5 mM HEPES; pH9.5 buffer: 50 mM NaCl, 10 mM CHES). After the final wash, the supernatant was removed, and the sample resuspended in 100 µL of the above‐described buffer. The samples were then placed in a quartz cuvette (Thorlabs Inc., Newton, NJ). UV‐Vis‐NIR absorption measurements were performed at room temperature on a Cary 6000i instrument equipped with a diffuse reflectance accessory (DRA) (Agilent Technologies, Santa Clara, CA). The samples were scanned at a rate of 600 nm min−1 over the range of 200‐900 nm.
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!