Blood monocytes isolated from HdhQ150 mice were seeded at 100,000 cells into 96-well plates in 100 μl R10 media. Due to low yields from R6/2 mice, cells were pooled according to genotype and multiple wells were seeded at 100,000 cells into 96-well plates in 100 μl R10 media. Splenic and bone marrow monocytes were seeded in 24-well plates at a density of 500,000 cells per well. All cultures were primed with 10 ng/ml murine IFNγ and stimulated with 2 μg/ml LPS. After 24 h, supernatants were collected for analysis of cytokine levels using multiplex ELISAs (mouse pro-inflammatory 7-Plex tissue culture kit measuring IFNγ, IL-1β, IL-10, IL-12p70, IL-6, mKC, TNFα) according to the manufacturer's instructions (Meso Scale Discovery). Cells were lysed in RIPA buffer (25 mM Tris–HCl pH 7.6, 150 mM NaCl, 1% NP-40, 1% sodium deoxycholate, 0.1% sodium dodecyl sulphate (SDS)) for protein measurement by BCA assay (Thermo Scientific).
Mouse pro inflammatory 7 plex tissue culture kit
Manufactured by Mesoscale
Sourced in United States
The Mouse pro-inflammatory 7-Plex tissue culture kit is a multiplex assay designed to quantify the levels of seven mouse pro-inflammatory cytokines and chemokines in tissue culture samples. The kit utilizes Mesoscale's electrochemiluminescence detection technology to provide sensitive and accurate measurements of these analytes.
Automatically generated - may contain errors
3 protocols using mouse pro inflammatory 7 plex tissue culture kit
1
Cytokine Profiling of Monocytes from Huntington's Disease Mice
2
Quantification of Cytokines and Interferons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokine quantities for IL-6, TNF-a, IL-10, CXCL1, IL-12p40, and IFN-g in the lung tissue were determined by the mouse proinflammatory 7-plex tissue culture kit (Meso Scale Discovery, cat K15012B-1) using 25ul of homogenized lung tissue according to manufacturer’s instructions. The concentration of IFN-α and IFN-β from mouse cell supernatant and mouse lung tissue was measured with VeriKine mouse IFN-α ELISA kit (PBL, cat 42400) and VeriKine mouse IFN-β ELISA kit (PBL, cat 42400), respectively.
The mRNA level of TNF-α, IFN-β, IL-6, and IFN-α in GM-DCs were tested with SYBR green method as described above. Primers used for the cytokine test were the following:
TNF-α-F, 5'-GGCAGGTCTACTTTGGAGTCATTG-3’, TNF-α-R, 5'-ACATTCGAGGCTCCAGTGAATTCGG-3’; IFN-β-F, 5'- AAGAGTTACACTGCCTTTGCCATC-3’, IFN-β-R, 5'- CACTGTCTGCTGGTGGAGTTCATC-3’; IL-6-F, 5'-GACAAAGCCAGAGTCCTTCAG AGAG-3’, IL-6-R, 5'-CTAGGTTTGCCGAGTAGATCTC-3’; IFN-α-F, 5'-CGCAGGAGAAGGTGGATGCCCAG-3’, IFN-α-R, 5'-CAGCACATTGGCAGAGGAAGACAGG-3’ [19 (link)].
The mRNA level of TNF-α, IFN-β, IL-6, and IFN-α in GM-DCs were tested with SYBR green method as described above. Primers used for the cytokine test were the following:
TNF-α-F, 5'-GGCAGGTCTACTTTGGAGTCATTG-3’, TNF-α-R, 5'-ACATTCGAGGCTCCAGTGAATTCGG-3’; IFN-β-F, 5'- AAGAGTTACACTGCCTTTGCCATC-3’, IFN-β-R, 5'- CACTGTCTGCTGGTGGAGTTCATC-3’; IL-6-F, 5'-GACAAAGCCAGAGTCCTTCAG AGAG-3’, IL-6-R, 5'-CTAGGTTTGCCGAGTAGATCTC-3’; IFN-α-F, 5'-CGCAGGAGAAGGTGGATGCCCAG-3’, IFN-α-R, 5'-CAGCACATTGGCAGAGGAAGACAGG-3’ [19 (link)].
3
Cytokine Profiling in Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pro-inflammatory cytokines were analyzed from culture media post 48-hour culture. Media was assessed for mouse tumor necrosis factor (TNF)-α, KC-GRO, interferon (IFN)-γ, interleukin (IL)-1β, IL-10, IL-6 and IL-12p70 using a Mouse ProInflammatory 7-Plex Tissue Culture Kit (Meso Scale Diagnostics, Rockville, Maryland, USA). Human IL-1β, IL-12p70, IL-6, IL-8, IL-10, IL-13, TNF-α and IFN-γ cytokine analysis was demonstrated using a Human ProInflammatory Panel kit (Meso Scale Diagnostics, Rockville, Maryland, USA). The plates were loaded into an MSD-SECTOR® instrument for electrochemiluminescence analysis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!