Mouse Cytokine Array Profiling
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Corresponding Organization :
Other organizations : Massachusetts Institute of Technology, Joslin Diabetes Center, University of Illinois at Chicago, Boston Children's Hospital, Harvard University, NanoScale Corporation (United States), University of Massachusetts Chan Medical School
Variable analysis
- Proteome Profiler Mouse Cytokine Array Panel A kit (Cat#ARY006, R&D system)
- Antibody-antigen complexes visualized using Odyssey Detection (Li-Cor, Serial No. ODY-2329) at 800 nm wavelengths
- Densities of the spots analyzed by Image J software
- 200 µl of protein solution
- 100 µl of sample buffer (array buffer 4)
- 1.2 ml of block buffer (array buffer 6)
- 15 µl of reconstituted Mouse Cytokine Array Panel A Detection Antibody Cocktail
- Incubation at room temperature for 1 h
- Incubation of array membrane with block buffer (array buffer 6) for 2 h on a rocking platform shaker
- Incubation of prepared sample/antibody mixture on the membrane overnight at 4 °C on a rocking platform shaker
- Washing with 20 ml of 1× wash buffer for 10 min on a rocking platform shaker for three times
- Rinsing with deionized water once
- Probing with Fluorophore-conjugated streptavidin (1:5,000 dilution, Cat#926-32230, Li-Cor) at room temperature for 30 min on a rocking platform shaker
- Washing with wash buffer for three times
- Rinsing with deionized water once again
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