Cells were plated in triplicate in 6-well plates and treated for 3 days with or without 1 μg/mL doxycycline hydrochloride (Thermo Fisher Scientific, Cat# BP2653-5) in 2 mL media. Conditioned media was collected and aliquots (75 μL) were then analyzed using the Mouse Cytokine/Chemokine 44-Plex Discovery Assay® Array from Eve Technologies.
Mouse cytokine chemokine 44 plex discovery assay array
Manufactured by Eve Technologies
The Mouse Cytokine/Chemokine 44-Plex Discovery Assay® Array is a multiplex assay designed to simultaneously quantify the levels of 44 different mouse cytokines and chemokines in a single sample. The assay utilizes Luminex® technology to provide a sensitive and efficient way to measure these important signaling molecules.
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2 protocols using mouse cytokine chemokine 44 plex discovery assay array
1
Cytokine/Chemokine Profiling of Doxycycline-Treated Cells
2
Multiplex Cytokine Analysis of Intestine
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Small (~5 mm) pieces of small intestine were homogenized for 30 s on ice with a disposable mini-pestle (Pellet Pestle Cordless Motor, Fisher Scientific) in 1.5 ml microfuge tubes using RIPA extraction buffer (Cell Signaling Technology, Cat No 9806) and a cocktail of protease inhibitors (HALT, Thermo Scientific, Cat No 78429). After centrifugation (20,000 x g, 4°C, 15 min) to remove tissue debris, supernatants were recovered and measured for protein content (BCA Protein Assay, Fisher Scientific). Samples were diluted to 1 mg/ml protein with extraction buffer, and submitted for multiplex cytokine analysis (Mouse Cytokine/Chemokine 44-Plex Discovery Assay Array, Eve Technologies).
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