Absolute levels of IL-16, IL-27, IL-1α, IL-1β and IL-6 in lysates prepared from human eosinophils and IL-16 in mouse eosinophils was determined using human DuoSet ELISAs from R&D Systems, including DY316 for IL-16; 15.6–1000 pg/mL range, DY2526 for IL-27; 156 – 10,000 pg/mL range; DY206 for IL-6, 9.38 – 600 pg/mL range; DY200 for IL-1α, 7.8 – 500 pg/mL range, and DY201 for IL-1β, 3.91 – 250 pg/mL range and mouse DuoSet ELISA DY1727 for IL-16; 46.9–3000 pg/mL range) respectively.
Human duoset elisas
Manufactured by R&D Systems
Sourced in United States
Human DuoSet® ELISAs are immunoassay development kits that contain the necessary components to develop quantitative sandwich ELISA for the measurement of human proteins in biological samples.
Automatically generated - may contain errors
4 protocols using human duoset elisas
1
Eosinophil Cytokine Quantification
2
Cytokine Quantification in Vitro
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human DuoSet® ELISAs (R&D Systems, Minneapolis, Minnesota, USA) were used to estimate the protein concentrations of CCL2, CXCL10, CXCL1, CXCL8, CXCL10, and CXCL11 chemokines in HIO supernatants, according to the manufacturer's instructions. Briefly, flat 96-well plates were coated overnight with a capture antibody for each chemokine, and the following day, plates were incubated with the recommended blocking buffer for 2 h. Next, duplicates of each supernatant and known concentrations of chemokine samples were added in wells and incubated for 2 h, and then, a biotinylated detection antibody for each chemokine was added for another 2 h. Streptavidin-horseradish peroxidase was then added for 20 min, and the following addition of tetramethylbenzidine with H2O2 produced different optical densities (OD) of color which were measured at 450 nm on a microplate reader (DIAReader ELX800; DIALAB, Wr. Neudorf, Austria). The chemokine concentration was calculated using a linear standard curve according to the manufacturer's instructions.
3
Quantifying Chemokine Levels in hPSMs and A549 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human DuoSet® ELISAs (R&D Systems, Minneapolis, MN, USA) were used to estimate the protein concentrations of CCL2, CCL10, CXCL1, CXCL8, CXCL10 and CXCL11 chemokines in hPSMs and A549 cells’ supernatants, according to the manufacturer’s instructions and as previously described [61 (link)]. Briefly, flat 96-well plates were coated overnight with capture antibody for each chemokine, and the following day, plates were incubated with the recommended blocking buffer for 2 h. Next, duplicates of each supernatant and known concentrations of chemokine samples were added in wells, incubated for 2 h, and then, biotinylated detection antibody for each chemokine was added for another 2 h. Streptavidin-horseradish peroxidase was then added for 20 min and the following addition of tetramethylbenzidine with H2O2 produced different optical densities (OD) of color which were measured at 450 nm on a microplate reader (Diareader EL×800; Dialab, Wr. Neudorf, Austria). The chemokines concentration was calculated using a linear standard curve according to the manufacturer’s instructions.
4
Quantification of IL6 and IL8 using ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IL6 and IL8 levels were quantified using R&D Systems human duoset ELISAs with a modified europium protocol. Briefly, black nunc plates were coated with capture antibody in coating buffer and incubated overnight at 4 °C. Plates were blocked with PBS/3% BSA before samples and standards added and incubated for 2 h. Detection antibody was then added for 2 h before Streptavidin-Europium diluted 1:1000 in DELFIA® assay buffer (both Perkin Elmer) was added for 30 min. Finally, DELFIA® enhancement solution (Perkin Elmer) was added for 10 min before reading on an Envision 2103 Multilabel Reader (Perkin Elmer). Plates were washed in PBS/Tween20 between each of these steps.
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!