The cell proliferation assay was used to analyze the proliferation of endothelial cells following the manufacturer’s protocols (Cell Signaling Technology). The H9-ECs were incubated in EGM-2 medium along with BrdU solution for 6 hours at 37 °C. HRP conjugate substrate was subsequently added and the absorbance was read at 450 nm by MD M5 SpectraMax (Molecular Devices). All experiments were performed in triplicates and data were analyzed using GraphPad Prism 6.
Md spectramax m5
Manufactured by Molecular Devices
Sourced in United States
The MD SpectraMax M5 is a multi-mode microplate reader that can perform absorbance, fluorescence, and luminescence measurements. It is designed to support a wide range of applications in life science research and drug discovery.
Automatically generated - may contain errors
Lab products found in correlation
Mts assay kit, by Promega (3 mentions)
Cell proliferation assay, by Cell Signaling Technology (2 mentions)
Mtt cell proliferation viability assay kit, by R&D Systems (2 mentions)
Prism 6, by GraphPad (1 mentions)
Microplate reader, by Agilent Technologies (1 mentions)
Mtt solution, by Wuhan Servicebio Technology (1 mentions)
Cell proliferation kit, by Merck Group (1 mentions)
Radioimmunoprecipitation assay (ripa), by Cell Signaling Technology (1 mentions)
Catalog 911 mp, by R&D Systems (1 mentions)
Genequant 1300, by GE Healthcare (1 mentions)
Methylflash methylated dna quantification kit, by Epigentek (1 mentions)
Qiaamp dna mini kit, by Qiagen (1 mentions)
Double antibody sandwich elisa, by Elabscience (1 mentions)
19 protocols using md spectramax m5
1
Endothelial Cell Proliferation Assay
2
Evaluating iPSC-EC Proliferation
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A cell proliferation assay was used to analyze the proliferation of ECs following the manufacturer's protocols (Cell Signaling Technology). The iPSC-ECs were incubated in EGM-2 medium along with bromodeoxyuridine solution for 6 h at 37°C. Horseradish peroxidase conjugate substrate was subsequently added, and the absorbance was read at 450 nm by MD M5 SpectraMax (Molecular Devices). All experiments were performed in triplicates, and data were analyzed using GraphPad Prism 6 (GraphPad Software).
3
Serum and Urine Mineral Analysis
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The concentrations of Ca and Cr in the serum and urine of the mice were measured using standard colorimetric methods with a microplate reader (Bio-Tek, Winooski, VT, USA). Serum was collected by cardiac exsanguination under light ether anesthesia. After 24 h urine was collected by metabolic cages. The level of Ca in the urine was corrected by the concentration of urine Cr levels. The serum levels of tartrate-resistant acid phosphatase-5b (TRACP-5b) and osteocalcin (OCN) were detected using a mouse bioactive enzyme-linked immunosorbent assay (ELISA; Immutopics, Inc., San Clemente, CA, USA) with an ELISA reader (MD SpectraMax M5; Molecular Devices).
4
Measuring NSC-34 and HT22 Proliferation
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The proliferation of NSC-34 and HT22 was examined using an MTS assay kit (Promega Corporation, USA). The optical density was observed with an enzyme-linked immunosorbent assay (ELISA) reader (MD SpectraMax M5; Molecular Devices, LLC, USA) at 492 nm.
5
Caspase-3 Activity Assay
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Cells (2×106 cells/well) lysates were prepared in NP-40 buffer at ice-bath condition for 15 min and centrifuged at 900 × g for 10 min at 4°C, and the supernatant was collected. In brief, 20 µl of cell lysate incubated with anti-caspase-3 antibody (cat. no. sc-7272; dilution, 1:200) at 37°C for 1 h. The immunocomplexes were then incubated with peptide substrate (2 µl of 10 mM acetyl-Asp-Glu-Val-Asp-p-nitroanilide) in assay buffer (100 mM Hepes, pH 7.5, 20% v/v glycerol, 5 mM dithiothreitol, and 0.5 mM EDTA) for 2 h at 37°C. The release of p-nitroaniline was measured at 405 nm using an ELISA reader (MD SpectraMax M5; Molecular Devices, LLC, Sunnyvale, CA, USA) according to the manufacturer's protocol.
6
Antibody Titer Quantification by ELISA
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The titers of vaccine-induced antibodies in mouse sera against Aβ42 peptide were determined by indirect enzyme-linked immunosorbent assay (ELISA). Briefly, 96-well ELISA plates were coated with Aβ42 peptide (0.5 μg/well) at 4 °C overnight and blocked with 3% (w/v) bovine serum albumin in PBS for 2 h at 37 °C. After blocking, 2-fold serial dilution of sera were added in triplicates and incubated for 2 h at 37 °C. The bound serum antibodies were detected with HRP-conjugated anti-mouse IgG and TMB substrate. The reaction was stopped by adding 2 M H2SO4, and the absorbance of OD450 nm were read on microplate reader MD-SpectraMax M5 (Molecular Device, Sunnyvale, CA, USA).
The levels of different IgG isotypes were also assayed by ELISA with the HRP-conjugated anti-mouse -IgG1, IgG3, -IgG2a and -IgG2b antibodies as the secondary antibodies (Abcam, Cambridge, UK), respectively.
The levels of different IgG isotypes were also assayed by ELISA with the HRP-conjugated anti-mouse -IgG1, IgG3, -IgG2a and -IgG2b antibodies as the secondary antibodies (Abcam, Cambridge, UK), respectively.
7
MTT Assay for Lung Cancer Cells
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Lung cancer cells with or without CES2 knockdown were seeded in 96-well plates (5 × 103 cells/well) and incubated overnight. Next, the cells were treated with GDNT or MMF at different concentrations, or co-treated with 6.25 μM GDNT and 5 μg/ml MMF at 37°C for 24 h. Thereafter, 20 μl MTT solution (G4101-1, Servicebio, China) was added into each well for further 4-h incubation. Following formazan dissolution by DMSO, MD SpectraMax M5 microplate reader (Molecular Devices, USA) was utilized to detect cell absorbance at 570 nm.
8
High Glucose-Induced Osteoblast MTT Assay
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The osteoblasts were cultured in 96-well plates (at 103 cells/well). An MTT assay was conducted after 1, 4 or 7 days of exposure to high glucose (16.5 mM) using a cell proliferation kit (Sigma, Oakville, ON, Canada) according to the manufacturer's instructions. After 24 h, the culture medium was replaced with 100 ml of MTT (0.5 mg/ml). The black crystals that formed after 2–3 h were dissolved with acidified isopropanol and the absorbance was measured at 570 nm using a microplate reader (MD SpectraMax M5; Molecular Devices, Sunnyvale, CA, USA).
9
Cytokine Profiling in Mouse Blood
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At various time-points, blood samples (approximately 1.2 mL each mouse) were separated from heart blood and centrifuged at 1200 g for 15 min at 4°C. The levels of cytokines, tumor necrosis factor-alpha (TNF-α, Cat. No: E-EL-M0049c), interleukin-1beta (IL-1β, Cat. No: E-EL-M0037c), and interleukin-18 (IL-18, Cat. No: E-EL-M0730c) (Elabscience Biotechnology Co., Ltd, China) were assayed using a double antibody sandwich ELISA following the manufacturer's instructions and evaluated using an ELISA reader (MD SpectraMax M5; Molecular Devices, USA).
10
Caspase-3 Activity Measurement
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HSFBs lysates were prepared and incubated with anti-caspase 3. Immunocomplexes were incubated with peptide substrate in assay buffer for 2 h at 37°C. Release of p-nitroaniline was measured at 405 nm using an ELISA reader (MD SpectraMax M5; Molecular Devices, Sunnyvale, CA, USA) according to the manufacturer's protocol.
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