The control HIBECs and rCsGSTos-absorbed HIBECs were incubated with IHBEC-1 medium for 24 h at 37 °C in a 5% CO2 incubator in the presence or absence of CHP (50–300 μM). The cells were also exposed to CHP (150 μM) for different time intervals from 30 min to 72 h, and cell viability was spectrophotometrically detected using a D-plus CCK cell viability assay kit (Donginbiotech, Seoul, Korea) at 450 nm (NEO microplate reader, Biotek, Winooski, VT, USA). HIBECs incorporated with heat-inactivated (95 °C for 10 min) rCsGSTos for 1 h (50 µg/mL) were used as controls.
Neo microplate reader
Manufactured by Agilent Technologies
Sourced in United States
The NEO microplate reader is a compact and versatile instrument designed for high-performance absorbance and fluorescence measurements in microtiter plates. It offers accurate and reliable data acquisition for a wide range of applications, including cell-based assays, enzyme kinetics, and ELISA.
Automatically generated - may contain errors
7 protocols using neo microplate reader
1
HIBEC Viability under CHP Treatment
2
Luminescence Spectroscopy of Lanthanide-CPA Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Excitation and emission spectra of the CPA/lanthanide complexes were determined using a Neo microplate reader (BioTek, Winooski, VT, USA). All spectra were collected at 25 °C in MeOH/H2O (9 + 1) using normal (not time resolved) luminescence. Test volumes were 300 µL. Experiments were conducted in black microtiter plates (Corning, Inc., Kennebunk, ME, USA). General settings for the instrument included a gain setting of 150, scanning in 1 nm increments, with 50 measurements per increment. Lamp energy was set to “low”, and distance of the optics above the microplate was set to 4.5 mm. The read speed was “normal”. Emission scans were collected over the range of 500 to 650 nm at an excitation of 290 nm. Excitation scans were collected over the range of 250 nm to 360 nm with monitoring emission at either 545 nm (Tb3+) or 615 nm (Eu3+). To demonstrate the enhancement, excitation and emission spectra were collected with and without CPA. The concentrations that were used were determined empirically and were selected to use as little of the reagents as possible while still yielding spectra with significant signal (5000 counts per second or higher) to allow for direct comparisons of sensitivity. For Eu3+ this was with EuCl3 at 2.5 µM, with and without CPA at 25 µM. For Tb3+ this was with TbCl3 at 0.25 µM with and without CPA at 1.5 µM.
3
ELISA for Anti-EgDM9 Antibody Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After a checkerboard titration, 100 μl of EgDM9 (2.5 μg/ml suspended in 100 mM carbonate-bicarbonate buffer, pH 9.6) was coated to the wells of a flat-bottom 96-well microplate (Greiner Bio-One, Kremsmuenster, Austria) overnight at 4°C. One hundred microliters of serum samples (1:200 dilution in phosphate buffered saline containing 0.05% Tween 20 [PBS/T]) were incubated for 2 hr at 37°C, subsequently 1:2,000 diluted HRP-conjugated anti-human IgG antibody (100 μl, Cappel) was incubated for 2 hr at 37°C. Color reactions were developed with 100 μl of 1% o-phenylenediamine (Sigma-Aldrich) supplemented with 0.03% H2O2 for 20 min in the dark. Reactions were stopped using 2 N H2SO4 (50 μl, Sigma-Aldrich). The absorbance was measured at 450 nm on a NEO microplate reader (Biotek, Winooski, Vermont, USA). Sera from healthy donors and PBS/T were used as negative and blank controls, respectively. Results were determined after correction with appropriate blank.
4
Checkerboard ELISA for rEgAgB IgG
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After a checkerboard titration, 100 µl of each rEgAgB (1.5 µg/ml suspended in 100 mM carbonate-bicarbonate buffer, pH 9.6) was used to coat the wells of a flat-bottom 96-well microplate (Greiner Bio-One, Kremsmuenster, Austria) overnight at 4 °C. One hundred microliters of serum samples diluted 1:100 in phosphate buffered saline containing 0.05% Tween 20 (PBS/T) were incubated at 37 °C for 2 h, after which 100 µl of horseradish peroxidase-conjugated anti-human IgG antibody (1:4000 dilution in PBS/T; MP Biochemicals, Santa Ana, CA, USA) was further incubated at 37 °C for 2 h. Color reactions were developed with 100 µl of 1% o-phenylenediamine (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 0.03% H2O2 for 20 min in the dark. Reactions were halted by adding 50 µl of 2 N H2SO4 (Sigma-Aldrich). The absorbance was measured at 450 nm on an NEO microplate reader (Biotek, Winooski, VT, USA). Sera from healthy donors and PBS/T were used as negative and blank controls, respectively. All results were determined after correction with appropriate blank. Each sample was independently assayed in triplicate.
5
Serological ELISA Assay for PwAWE and rPwYF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After a checkerboard titration, 100 μl of PwAWE and rPwYF (2.5 μg/ml each) suspended in 100 mM carbonate-bicarbonate buffer (pH 9.6) were coated to the wells of a flat-bottom 96-well microplate (Greiner Bio-One, Kremsmünster Austria) overnight at 4°C. Sera diluted to 1:200 in PBS containing 0.05% Tween 20 (PBS/T) or undiluted CSFs (100 μl each) were incubated for 2 h, subsequently with 1:2,000 diluted horseradish peroxidase (HRP)-conjugated anti-human IgG antibody (100 μl; heavy- and light-chain specific, MP Biochemicals, Santa Ana, CA, USA) for 2 h. The color reaction was developed with 1% o-phenylenediamine (100 μl; Sigma-Aldrich) supplemented with 0.03% H2O2 for 20 min in the dark. The reaction was stopped using 2 N H2SO4 (50 μl; Sigma-Aldrich). The absorbance (abs.) was measured at 450 nm on a NEO microplate reader (Biotek, Winooski, VT, USA). All results were determined after correction with PBS/T blank.
6
Fluorescence Analysis of Antibody-Dye Interactions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Preliminary experiments indicated that the mAbs could influence the fluorescence of CTV in aqueous buffer. To examine this further, mixtures of mAb or BSA were combined with CTV in aqueous buffer (10 mM PBS, pH 7.2). The final concentrations of mAb (or BSA) and CTV were 2.0 µM and 1.25 µM, respectively. The volumes of test solutions were 0.32 mL. Excitation and emission spectra were collected using a Neo microplate reader (BioTek, Winooski, VT, USA). Emission scans were collected using an excitation of 420 nm, with emission monitored over the range of 450 to 700 nm in 1 nm increments. Excitation scans were collected using an emission of 570 nm, with excitation provided over the range of 360 to 500 nm in 1 nm increments. Both excitation and emission data were collected with the top optics of the instrument and the following parameters: gain 150, normal reading speed, optics positioned 4.50 mm above the sample, and temperature 20.5 °C. Data from triplicate plates were averaged and imported into TableCurve. Spectra were smoothed using Savitzky–Golay filtering.
7
ELISA for Detecting Antibodies to rCsGSTo1/2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After checkerboard titration, 100 mL of rCsGSTo1 or rCsGSTo2 protein (each 1.5 mg/mL in 100 mM carbonate-bicarbonate buffer, pH 9.6) was coated onto the wells of a flat-bottom 96-well microplate (Greiner Bio-One, Austria) overnight at 4 C. Serum samples (100 mL) were diluted at 1:100 in phosphate-buffered saline containing 0.05% Tween 20 (PBS/T) and incubated for 2 h at 37 C. Horseradish peroxidase-conjugated goat anti-human IgG antibody (Cappel, USA) diluted at 1:1000 (100 mL) was incubated for 2 h at 37 C. Colour reaction was developed with 100 mL of 1% o-phenylenediamine (Sigma-Aldrich) containing 0.03% H 2 O 2 for 20 min in the dark and stopped by adding 50 mL of 2 N H 2 SO 4 (Sigma-Aldrich).
Absorbance was measured at 450 nm (NEO microplate reader; Biotek, USA). Sera of healthy persons and PBS/T were used as negative and blank controls, respectively. All results were measured after appropriate blank correction. Each sample was assayed in triplicate.
Absorbance was measured at 450 nm (NEO microplate reader; Biotek, USA). Sera of healthy persons and PBS/T were used as negative and blank controls, respectively. All results were measured after appropriate blank correction. Each sample was assayed in triplicate.
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!