To measure intracellular ROS or NO level, NP cells was incubated with DCFH-DA (10 μM) for 30 min or with DAF-FM DA (5 μM) for 20 min in dark at 37°C. After washed with PBS 3 times, NP cells were collected and suspended in fresh medium. The fluorescence intensity was detected using a microplate reader (Omega Bio-Tek, Inc.). The excitation wavelengths for DCFH-DA and DAF-FM DA are 488 and 495 nm respectively. The experiments were repeated 3 times.
Microplate reader
Manufactured by Omega Bio-Tek
Sourced in United States
The Microplate reader is a laboratory instrument used to measure the absorbance, luminescence, or fluorescence of samples in a microplate format. It is designed to accurately detect and quantify various biological, chemical, or physical properties of samples contained in the microplate wells. The core function of the Microplate reader is to provide precise and reproducible measurements for a wide range of applications in life science research, drug discovery, and clinical diagnostics.
Automatically generated - may contain errors
Lab products found in correlation
Mtt solution, by Merck Group (2 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (2 mentions)
Sod assay kit, by Beyotime (1 mentions)
Hrp conjugated goat anti mouse igg antibody, by Jackson ImmunoResearch (1 mentions)
Tmb solution, by Solarbio (1 mentions)
Cck 8 solution, by Beyotime (1 mentions)
Cck 8 kit, by Beyotime (1 mentions)
Pd98059, by Merck Group (1 mentions)
Mts reagent kit, by Promega (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Alp assay kit, by Beyotime (1 mentions)
Il 1β, by Boster Bio (1 mentions)
Icam 1, by Boster Bio (1 mentions)
Tnf α, by Boster Bio (1 mentions)
Interleukin 6 (il 6), by Boster Bio (1 mentions)
Malondialdehyde assay kit, by Nanjing Jiancheng (1 mentions)
Cell counting kit 8 cck 8 reagent, by Dojindo Laboratories (1 mentions)
Mtt assay, by Avantor (1 mentions)
Cell counting kit 8 (cck8), by Merck Group (1 mentions)
U0126, by Merck Group (1 mentions)
28 protocols using microplate reader
1
Intracellular ROS and NO Detection
2
Measuring Cellular SOD Activity
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SOD activity was determined using an SOD assay kit (Beyotime Institute of Biotechnology). In brief, SOD sample preparation solution was used to lyse PC12 cells, and the solution was centrifugated for 5 min at 4˚C (12,000 x g) to collect cell supernatant. Corresponding solution was added for 30 min at 37˚C and the absorbance value was recorded using a microplate reader (Omega Bio-Tek, Inc.) at 450 nm wavelength.
3
MTT Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following the treatment of cells, 10 µL of MTT solution (5 mg/mL) was added to each well of the 96-well plate. Following incubation for 4 h, the medium was removed and 150 µL of dimethyl sulfoxide (DMSO) was added. The absorbance value at 570 nm was read using a microplate reader (Omega Bio-Tek, Inc., Norcross, GA, USA).
4
MTT Cell Viability Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CaSki and HeLa cells were harvested, seeded in 96-well plates (1×103 cells/ml per well) and grown until 80% confluence. MTT solution (5 mg/ml; Sigma-Aldrich; Merck KGaA) was subsequently added to the cells (15 µl/well). Following 4 h of incubation of the cells with MTT solution at 37°C, DMSO was added (150 µl/well) to dissolve the formazan crystals. A microplate reader (Omega Bio-Tek, Inc.) was used for spectrophotometry-based measurements of the optical density at 490 nm.
5
Conformational Antibody Assay for VP2 Capsids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An indirect enzyme-linked immunosorbent assay was carried out using the conformational antibody 10A9, which only recognizes assembled capsids, as prepared in our previous study. First, 96-well plates were coated with puri ed mutant VP2 proteins, wild-type VP2 protein, or pET 28a empty vector, and then blocked with 5% skim milk in PBST at 4°C at least 8 hours. The conformational antibody 10A9 was used as the primary antibody and added into 96-well plates at 37°C for 30 minutes. After washing with PBST for ve times, an HRP-conjugated goat anti-mouse IgG antibody (Jackson ImmunoResearch, Philadelphia, PA, USA) was used as the secondary antibody and incubated in the plate at 37°C for 45 minutes. Finally, antibody binding was analyzed by adding TMB solution (Solarbio, Beijing, China), and 2 M H 2 SO 4 was added to stop the reaction. The optical density at 450 nm was recorded using a microplate reader (Omega Bio-Tek, Inc., Norcross, GA, Germany). All the experiments were performed in triplicate.
6
Cell Proliferation Assay with CCK-8
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation was analyzed using a Cell Counting Kit-8 (CCK-8; Dojindo Molecular Technologies, Inc.). HUVECs were seeded into 96-well plates at a density of 2x103 cells/well and cultured at 37˚C with 5% CO2. CCK-8 reagent was added to the cells at time points of 0, 24, 48 and 72 h with a further 2-h incubation. The absorbance was measured at a wavelength of 450 nm using a microplate reader (Omega Bio-Tek, Inc.).
7
TGF-β1-induced cell proliferation assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HaCaT cells were seeded in 96-well plates at a density of 2×103 cells/well and treated with TGF-β1 (10 ng/mL), either with or without Ang-(1-7) (1 μM) for 48 h. Then, 10 μL of CCK-8 solution were added to each well and incubated at 37°C for 2 h, according to the manufacturer’s protocol (Beyotime Institute of Biotechnology). The optical density of the cells was measured at a wavelength of 450 nm using a microplate reader (Omega Bio-Tek, Inc.).
8
Colorimetric Assay for Cell Viability
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW480 cells were plated in 96-well plates (8×103 cell/well). Following the corresponding treatment, the survival rate of the cells was detected with the CCK-8 kit (Beyotime Institute of Biotechnology, Shanghai, China). Subsequently, 10 µL CCK-8 solution was added to each well and incubated for 2 h according to the manufacturer’s instructions. OD450 nm value was judged adopting a microplate reader (Omega Bio-Tek, Inc., Norcross, GA, USA).
9
Proliferation Assay of USP18-Knockdown HeLa Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Cell Counting Kit-8 (CCK-8; Wuhan Boster Biological Technology, Ltd.) assay was performed to assess the proliferation of HeLa cells following USP18-knockdown. Briefly, mock- and USP18-siRNA-transfected HeLa cells were seeded into 96-well plates at a density of 1.5×103 cells/well in a final volume of 100 µl RPMI-1640 medium supplemented with or without 80 µΜ of the ERK1/2 blocker PD98059 (Sigma-Aldrich; Merck KGaA). Following incubation at 37°C for 24, 48 or 72 h, 10 µl CCK-8 reagent was added to each well and incubated at 37°C for 1 h. Cell proliferation was subsequently analyzed at a wavelength of 450 nm, using a microplate reader (Omega Bio-Tek, Inc.). The inhibitory role of PD98059 on the proliferation of HeLa cells was estimated as follows: Inhibition rate (%)=(treatment with 0 µM PD98059-treatment with 80 µM PD98059)/treatment with 0 µM PD98059 ×100.
10
MTT Cytotoxicity Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
When cells reached 70% confluency in a 96-well microplate, they were cultured in a medium containing CDDP (0, 1, 5, 10, 20, 40 and 80 µM) or H2O2 (0, 5, 10, 25 and 50 µM) at 37°C for 24 h. Then, 10 µl MTT, dissolved in PBS, was added and the cells were incubated for 2 h at 37°C. The purple formazan was dissolved in PBS. The absorbance values of the cells were detected at 490 nm using a microplate reader (Omega Bio-Tek, Inc.). The IC50 was calculated with GraphPad Prism v.5.01 software (GraphPad software, Inc).
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!