Cell viability was determined using a cell counting kit (CCK)-8 assay (Catalogue No. KGA317; Nanjing KeyGEN BioTECH, Jiangsu, China). Briefly, 1 × 104 cells were seeded into 96-well plates one day prior to P. aeruginosa infection. Cells were then infected with P. aeruginosa, followed by addition of 10 µl CCK-8 reagent into the plate. Uninfected cells were used as controls. Absorbance was measured at 450 nm using a Bio-Rad ELISA plate reader (Bio-Rad Laboratories).
Elisa plate reader
Manufactured by Bio-Rad
Sourced in United States, Germany, Japan, India, Austria, Italy
The ELISA plate reader is a laboratory instrument designed to measure the absorbance of light in microtiter plates, commonly used for enzyme-linked immunosorbent assay (ELISA) experiments. It is capable of detecting and quantifying the presence of specific substances, such as proteins, antibodies, or other molecules, in liquid samples.
Automatically generated - may contain errors
Lab products found in correlation
Methyl thiazolyl tetrazolium (mtt), by Merck Group (7 mentions)
96 well microtiter plate, by Corning (5 mentions)
Dispenser washer, by Agilent Technologies (4 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Mtt assay, by Merck Group (3 mentions)
Penta his hrp conjugate, by Qiagen (3 mentions)
Gloxmax 20 20 luminometer, by Promega (3 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (3 mentions)
Axio observer z1, by Zeiss (3 mentions)
O phenylenediamine dihydrochloride, by Merck Group (2 mentions)
Il 23p40, by Elabscience (2 mentions)
Il 23p19, by Elabscience (2 mentions)
Ccl20, by Elabscience (2 mentions)
Matched antibodies, by R&D Systems (2 mentions)
Pipetting robot, by Agilent Technologies (2 mentions)
Celltiter 96 non radioactive cell proliferation assay, by Promega (2 mentions)
Ab100702, by Abcam (2 mentions)
Ab233621, by Abcam (2 mentions)
Dimethyl sulfoxide (dmso), by Beyotime (2 mentions)
96 well plate, by Corning (2 mentions)
244 protocols using elisa plate reader
1
Cell Viability Assay for P. aeruginosa Infection
2
Quantification of Plasma Aβ1-40 Levels
3
Caspase 3 Activity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Caspase 3 activity was estimated by using Ac-DEVD-pNA substrate (Santa Cruz Biotech, CA, USA). Tissue supernatant was first diluted in assay buffer (1X) containing 20 mM HEPES (pH 7.4), 0.1% CHAPS, 5 mM DTT and then DEVD-pNA substrate (2 mM) was added. The reaction plate was incubated at 37 °C overnight in a humidified incubator. Subsequently, contents of the plate were read at 405 nm in a BioRad Elisa plate reader.
4
Cytotoxicity Evaluation of Sealers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MTT assay (Sigma-Aldrich Co.) solution was prepared as 1 mg/mL in phosphate buffer saline just before use. 100-μL MTT dye was added to each well-containing cell treated with various extracts of sealers. Plates were incubated in a CO2 incubator for 3 h. Optical density was determined by eluting the dye, and the spectrophotometric absorbance was measured at 550 nm using a Bio-Rad ELISA plate reader (Alfred Nobel Drive, Hercules, CA, USA). The percentage of cell viability was calculated from the formula:
5
Cytotoxicity Evaluation of GLME in Cancer Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
PDL, Hek-293, MCF-7, and K-562 cell lines were used to test the cytotoxicity of GLME. The positive control was 5-Fluorouracil and carboplatin. After cultured cells in the T-75 flask, the cells were counted and seeded on 96-well plates with approximately 10,000 cells per well containing DMEM culture media and incubated to achieve 85 to 90% confluence. The previous media was then used to substitute 100 µL of GLME in a wide variety of concentrations. Next, in each well, 30 mL of MTT (3-(4,5 Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium) stock solution (concentration 4 mg mL-1) was transferred and incubated under normal conditions. Purple formazan crystals formed due to the viable cells' mitochondrial function, and we used 100 µL of dimethyl sulfoxide (DMSO) to dissolve these crystals. The plate was shaken in a double orbital manner (for 5 min) to dissolve formazan crystals completely. Finally, the optical absorption of the mentioned solution was recorded at 540 nm using an Elisa plate reader (Model 50, Bio-Rad Corp, Hercules, California, USA)33 (link),34 (link). All tests were accomplished in triplicate35 (link). The following equation describes the calculations of the cell viability:
6
Cell Viability Assessment of CuS-NO Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An MTT test was used to evaluate the effect of CuS–NO˙ NPs on cell viability. Briefly, HUVECs were inoculated in 96-well plates (3 × 103 cells per well). After 12 h, the culture medium supernatant was discarded. Then, the cells were washed with PBS (pH 7.4) and co-cultured with different concentrations of CuS–NO˙ NPs for 24 h. A 20 μL volume per well of MTT was added into 96-well plates. After 4 h, DMSO was used to dissolve the blue formazan crystals for 0.5 h. The absorbance of 96-well plates was obtained at 490 nm by an ELISA plate reader (BIO-RAD Inc., USA).
7
Bacterial Adhesion Quantification Methods
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A microscopic adhesion assay was used to assess bacterial adhesion on Cn substrates. Substrates were incubated for 2 h in 200-µl bacterial suspensions adjusted in PBS to a concentration of 109 cells ml−1. After 2 h, the substrates were gently rinsed by 3 consecutives washing in PBS and directly imaged using an inverted optical microscope (Zeiss Axio Observer Z1) equipped with a model C10600 Hamamatsu camera.
A crystal violet assay was also used. Microtiter wells were coated overnight at 4°C with 1 µg/well collagen type II–0.1 M sodium carbonate (pH 9.5). The plates were washed with 0.5% (vol/vol) PBS with Tween 20 (PBST). To block additional protein-binding sites, the wells were treated for 1 h at 22°C with 2% (vol/vol) bovine serum albumin (BSA)–phosphate-buffered saline (PBS). The wells were then incubated for 2 h at 37°C with 1 × 108S. aureus Phillips cells–PBS. After washing with PBS was performed, adhering cells were fixed with 2.5% formaldehyde for 30 min and stained with 1% crystal violet for 1 min. After washing, 100 µl of 10% acetic acid was added, and absorbance at 595 nm was recorded using an ELISA plate reader (BioRad).
A crystal violet assay was also used. Microtiter wells were coated overnight at 4°C with 1 µg/well collagen type II–0.1 M sodium carbonate (pH 9.5). The plates were washed with 0.5% (vol/vol) PBS with Tween 20 (PBST). To block additional protein-binding sites, the wells were treated for 1 h at 22°C with 2% (vol/vol) bovine serum albumin (BSA)–phosphate-buffered saline (PBS). The wells were then incubated for 2 h at 37°C with 1 × 108S. aureus Phillips cells–PBS. After washing with PBS was performed, adhering cells were fixed with 2.5% formaldehyde for 30 min and stained with 1% crystal violet for 1 min. After washing, 100 µl of 10% acetic acid was added, and absorbance at 595 nm was recorded using an ELISA plate reader (BioRad).
8
Mouse Forestomach Carcinoma Cell Line Study
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse forestomach carcinoma cell line (MFC) was purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai); aspirin was purchased from Sigma (St. Louis, MO, USA). RPMI-1640 medium (containing 15% fetal bovine serum, 100 µ/ml penicillin and 100 µ/ml streptomycin) were obtained from HyClone (Logan, UT, USA). Trypsin was obtained from Gibco (Grand Island, NY, USA). PBS was purchased from Shanghai Biological Engineering Company (Shanghai, China). MTT assay (5 mg/ml) was obtained from Sigma (St. Louis, MO, USA). Polyclonal rabbit anti-human E-cadherin was purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). FITC-labeled goat anti-rabbit IgG was from Beijing Zhong Shan Jinqiao Biological Technology Co., Ltd. (Beijing, China). Centrifuge was obtained from Thermo Fisher Scientific (Waltham, MA, USA) and syringes were from Bio-Rad (Berkeley, CA, USA). Inverted microscope was from Olympus (Tokyo Japan), Eppendorf tubes were obtained from Eppendorf (Hauppauge, NY, USA), super clean bench was from Thermo Fisher Scientific (Waltham, MA, USA), and cell counting chamber was from Shanghai Optical Instrument Factory (Shanghai, China). Paraffin slicing machine and tissue embedder were obtained from Leica (Mannheim, Germany) and ELISA plate reader was from Bio-Rad.
9
Cell Growth Determination by MTT Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cell growth rate was determined by MTT assay (Sigma-Aldrich). Briefly, cells (100 μl) at the logarithmic growth phase were seeded at a 1×104/ml density into 96-well culture plates. MTT solution (10 μl; 5 mg/ml) was added into each well and incubated at 37°C for 4 h. Following centrifugation at 1,409 × g for 10 min, the supernatant was discarded and 100 μl DMSO was added. When the remaining formazan pellet was dissolved completely, the absorbance values at a 570 nm wavelength were read on an ELISA plate reader (Bio-Rad, Hercules, CA, USA). The total procedure was repeated three times.
10
ELISA Binding Assay for 37LRP and APP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
High binding plates with 96 flat-bottomed wells (Corning, Amsterdam, Netherlands) were coated with GT1 and HeLa cell lysates, or BSA as a negative control, and incubated at 4 °C overnight. After a wash in PBS, residual binding sites were blocked for 1 h at 37 °C with 200 μL of blocking buffer (2% FCS, 1 mg/mL BSA, in PBS). Wells were incubated with 2 μg of pTrc-His B control eluate (diluted in PBS, 1 mg/mL BSA), or 2 μg of r37LRP (diluted in PBS, 1 mg/mL BSA), which both contained a 6 × His-tag, for 1 h at 37 °C. Each well was washed three times with wash buffer (0.5% Tween in PBS). Penta-His HRP conjugate (1:500) (Qiagen, Hilden, Germany) was added for 2 h at room temperature. After washing, substrate solution was added and absorbance was detected at 490 nm on an ELISA plate reader (Bio-Rad, Hercules, CA, USA). Binding affinity was determined by subtracting background absorbance (BSA wells).
For inhibition experiments, wells pre-coated with GT1 cell lysates were incubated with 2 μg of pTrc-His B, as control, and 2 μg of r37LRP, alone and in the presence of APP antibody (1:1000), or NSC48478 compound (20 μM).
For inhibition experiments, wells pre-coated with GT1 cell lysates were incubated with 2 μg of pTrc-His B, as control, and 2 μg of r37LRP, alone and in the presence of APP antibody (1:1000), or NSC48478 compound (20 μM).
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!