HBSS (with Ca2+ and Mg2+), Dulbecco’s Modified Eagle’s Medium (DMEM; high glucose), 0.05% trypsin solution and penicillin/streptomycin solution, and enzyme-free cell dissociation buffer were purchased from ThermoFisher Scientific (Waltham, MA, USA). Fetal bovine serum (FBS) was obtained from Bodinco (Alkmaar, the Netherlands). Linear 25 kDa polyethylenimine (PEI) was purchased from Polysciences (Warrington, PA, USA). Bovine serum albumin (BSA) was obtained from Melford (Ipswich, UK). Furimazine (N1130) was purchased from Promega (Madison, WI, USA). The 96-well white and black cell culture plates used were obtained from Greiner Bio-one (Kremsmünster, Austria). White low-volume 384-well plates were bought from Corning (Corning, NY, USA). Human recombinant CXCL12, CXCL10, and the fluorescently labeled chemokines CXCL12-AZD488, CXCL12-AZD546, CXCL12-AZD594, CXCL12-AZD647, and CXCL10-AZD488 were purchased from Protein Foundry (Milwaukee, WI, USA). IT1t (Bio-Techne Ireland Limited, Dublin, Ireland), AMD3100 and Burixafor were purchased from MedChemExpress (Princeton, NJ, USA), and VUF25444, VUF15485, VUF25550 and VUF16545 were synthetized in-house.
White low volume 384 well plates
Manufactured by Corning
Sourced in United States
The White low-volume 384-well plates are laboratory equipment designed for various scientific applications. The plates feature 384 individual wells, each with a low-volume capacity, providing a compact and efficient platform for conducting experiments and assays. The white color of the plates offers optimal light reflection and contrast, which can be beneficial for certain experimental setups. These plates are suitable for a range of laboratory procedures and are intended to be used in accordance with the specific requirements of the research or analysis being conducted.
Automatically generated - may contain errors
Lab products found in correlation
Enzyme free cell dissociation buffer, by Thermo Fisher Scientific (1 mentions)
Trypsin solution, by Thermo Fisher Scientific (1 mentions)
Hbss with ca2 and mg2, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
Amd3100, by MedChemExpress (1 mentions)
Linear 25 kda polyethylenimine pei, by Polysciences (1 mentions)
2 protocols using white low volume 384 well plates
1
Chemokine Receptor Binding Assay
2
Quantifying Myometrial ERK Phosphorylation
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Levels of phosphorylated ERK (pERK) were determined using the HTRF Cellul'ERK assay kit (Cisbio Bioassay, Codolet, France) as per the manufacturer's instructions.
Cell stimulation and lysis: Myometrial cells were seeded in 96-well plates at a density of 5000 cells/well and cultured overnight before serum starvation for 24 hours. Where appropriate, cells were pretreated with 100 ng/mL PTX for 18 hours. Cells were washed in m-KHB and equilibrated in buffer at 37°C for 30 minutes. Cells were challenged with compounds for time points indicated before aspiration and addition of 50 µL of supplied-lysis buffer (containing phosphatase inhibitors) for 30 minutes (room temperature).
Detection of pERK: 16 µL of each well was transferred in duplicate to white, low-volume 384-well plates (Corning®) for detection of pERK as per the Cellul'ERK assay manufacturer's instructions with fluorescence determined on a PHERstar FS plate reader.
Data analysis: Increases in pERK were calculated as % increase over basal (vehicle, unstimulated) cells.
Cell stimulation and lysis: Myometrial cells were seeded in 96-well plates at a density of 5000 cells/well and cultured overnight before serum starvation for 24 hours. Where appropriate, cells were pretreated with 100 ng/mL PTX for 18 hours. Cells were washed in m-KHB and equilibrated in buffer at 37°C for 30 minutes. Cells were challenged with compounds for time points indicated before aspiration and addition of 50 µL of supplied-lysis buffer (containing phosphatase inhibitors) for 30 minutes (room temperature).
Detection of pERK: 16 µL of each well was transferred in duplicate to white, low-volume 384-well plates (Corning®) for detection of pERK as per the Cellul'ERK assay manufacturer's instructions with fluorescence determined on a PHERstar FS plate reader.
Data analysis: Increases in pERK were calculated as % increase over basal (vehicle, unstimulated) cells.
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