Serum levels of IFN-γ, IL-2, IL-4, and IL-12 were determined by using a ProcartaPlex multiplex immunoassay panel (ThermoFisher, Waltham, MA, USA, cat. EPX110-20820-901) and Luminex 200 (Bio-Rad, Hercules, CA, USA). E0771 tumor samples were collected and snap frozen in liquid nitrogen. Samples were homogenized in lysis buffer (ThermoFisher, cat. EPX-99999-000) and centrifuged at 13,000 rpm for 10 min at 4 °C. Supernatant was collected and total protein concentration was normalized between all samples. Each serum sample was tested in two magnetic bead wells in mouse ProcartaPlex panels. The net mean fluorescence intensity (MFI) was averaged for three samples per treatment group for a biological replicate of n = 3/group and technical replicate of n = 6/group.
Procartaplex multiplex immunoassay panel
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ProcartaPlex multiplex immunoassay panel is a tool for simultaneous quantitative measurement of multiple analytes in a single sample. It enables efficient and accurate analysis of complex biological samples.
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Lab products found in correlation
2 protocols using procartaplex multiplex immunoassay panel
1
Multiplex Cytokine Analysis in Tumors
2
Microglial Secretome Effects on Photoreceptor Viability
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661W photoreceptor cells (gift from Dr. Muayyad Al-Ubaidi; University of Oklahoma, Oklahoma City, OK) were maintained in 96-well plates. Cultured retinal microglia from animals of different genotypes were allowed to condition in serum-free medium (DMEM/F12) for 48 h. Conditioned medium (100 µl) was then added to 661W cells for 16 h, and the resulting cell viability was assessed using an MTT cell assay kit (ATCC) following the manufacturer’s specifications. Protein analysis of conditioned medium was performed using a customized ProcartaPlex Multiplex Immunoassay panel (Thermo Fisher Scientific) according to the manufacturer’s instructions. In brief, magnetic beads were seeded in triplicate onto a 96-well plate, after which conditioned media (50 µl) from retinal microglia cultured from C57BL6/J, C3+/+; C3+/+.rd10, C3−/−.rd10, and CR3−/−.rd10 animals were added to each well and incubated overnight at 4°C. The beads were washed before the addition of the detecting antibodies followed by Streptavidin-PE addition and quantification on a Luminex 200 instrument (Luminex).
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