Murine cytokine concentrations were determined in serum samples isolated by cardiac puncture at the experimental end point. Cytokines were detected in duplicate wells per sample using the V-PLEX Proinflammatory Panel 1 Mouse Kit and V-PLEX Cytokine Panel 1 Mouse Kit (Meso Scale Diagnostics, LLC®) according to the manufacturer’s instructions. Data were analysed using DISCOVERY WORKBENCH® 4.0 software.
V plex cytokine panel 1 mouse kit
Manufactured by Mesoscale
The V-PLEX Cytokine Panel 1 Mouse Kit is a multiplex assay that allows for the quantitative measurement of multiple cytokines in mouse samples. The kit includes reagents and plates pre-coated with capture antibodies specific to the target cytokines.
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4 protocols using v plex cytokine panel 1 mouse kit
1
Murine Cytokine Profiling by ELISA
2
Multiplex Cytokine Profiling in Serum and Liver
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Meso Scale Discovery (MSD) multi-spot electrochemiluminescence assay system was used to quantify cytokine levels in blood serum or liver protein. V-PLEX Proinflammatory Panel 1 Mouse Kit (K15048D-1, Meso Scale Diagnostics) and V-PLEX Cytokine Panel 1 Mouse Kit (K15245D, Meso Scale Diagnostics) were used according to manufacturer’s instructions.
3
Cytokine Profiling of Brain Tumors
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Fifteen days after CT-2A tumor implantation, blood and brain tumors were collected from the CT-2A-bearing mice received RT + /- B-LNP/diABZI treatments as the treatment regimen described above. Protein was extracted from brain tumors following a previously published protocol51 (link). Cytokines were measured using a V-PLEX Cytokine Panel 1 Mouse Kit following the manufacturer protocol (Meso Scale Diagnostics).
4
Comprehensive Evaluation of Airway Inflammation
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BAL and evaluation of inflammatory cell infiltration were performed as described previously (82 (link)). Aliquots of cell-free BAL fluid were used to measure cytokines and chemokines via mesoscale technique using two different kits (V-Plex proinflammatory panel 1 mouse kit and V-Plex cytokine panel 1 mouse kit; MesoScaleDiscovery) according to manufacturer’s instructions. Total and ovalbumin-specific IgE were measured in serum samples by ELISA as described previously (84 (link)). IL-9 was measured in serum samples using mouse-IL-9 ELISA (BioLegend). For lung histology, after BAL, the lungs were excised and the left lobe fixed in 4% buffered formalin and embedded in paraffin. Sections of 4µm thickness were stained with hematoxylin-eosin (H&E) and periodic acid Schiff (PAS). Mucus hypersecretion and inflammatory cell infiltration were graded in a blinded fashion on a scale from 0 to 4 (0=none, 1=mild, 2=moderate, 3=marked, 4=severe), reflecting the degree of the pathological alteration (82 (link)).
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