Supernatants of vaginal washes were collected and tested for Interleukin-1ß (IL-1ß), TNF-α, IL-6 and IL-10 levels by specific ELISAs (Thermo Fisher Scientific). Cytokine titers were calculated relative to standard curves.
Specific elisa
Manufactured by Thermo Fisher Scientific
Specific ELISA is a laboratory equipment used for quantitative measurement of target analytes in a sample. It utilizes enzyme-linked immunosorbent assay (ELISA) technology to detect and measure the concentration of a specific substance in a solution.
Automatically generated - may contain errors
8 protocols using specific elisa
1
Vaginal Cytokine Profiling by ELISA
2
TRPC3 Inhibitor Modulates LPS-Induced Inflammation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
BALB/cJ mice were obtained from the University of Valladolid Animal House (10–12-week-old males). The mice were intraperitoneally injected with the TRPC3 specific inhibitor Pyr10 (1 mg/kg) (Sigma-Aldrich) for 1h and then treated with LPS at a sublethal dose of 5 mg/kg (i.p.). For the analysis of proinflammatory factors, animals were sacrificed by ketamine (100 mg/kg) / xylacine (10 mg/kg) administration and cervical dislocation 3 h after LPS treatment. Blood was collected through cardiac puncture. Livers were collected in RNAlater (Ambion) for further analysis by real-time PCR or in protein lysis buffer for western blot analysis. Serum from animals was used for quantification of TNF-α by specific ELISAs (Thermo) following the manufacturer’s instructions. BALB/cByJ- Lpin1fld/J mice carrying a spontaneous mutation in the Lpin1gene (fatty liver dystrophy, fld) were also used [33 (link)]. All procedures involving animals were carried out under the supervision of the Institutional Committee of Animal Care and Usage of the University of Valladolid (Approval No. 7406000), and are in accordance with the guidelines established by the Spanish Ministry of Agriculture, Food, and Environment, and the European Union.
3
Biomarkers Measured in Blood Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GDF‐15 was measured in ethylenediaminetetraacetic acid (EDTA) plasma with a specific enzyme‐linked immunosorbent assay (ELISA) (DY957, R&D Systems) according to the manufacturer´s manual. hsTnT and NT‐proBNP measurements were performed in heparin plasma using the Elecsys System (Roche Diagnostics). MR‐proANP, MR‐proADM, CT‐proET‐1 and copeptin were measured in EDTA plasma using specific sandwich immunoassays (BRAHMS). CRP and SAA levels were determined in EDTA and heparinized plasma by means of particle enhanced immunonephelometry using the BN II System (Siemens Healthcare Diagnostics). Serum IL‐6 was detected with a specific ELISA (eBioscience).
4
Modulation of T-cell Cytokine Production by PGE2
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1 × 106 FACS-sorted PDE4A- or control vector-transduced CD4+ and CD8+ T-cells were activated with anti-CD3/anti-CD28 coated microbeads (beads to cell ratio 1:2) in the presence or absence of the indicated concentrations of PGE2. After 24 and 72 h supernatants were harvested and levels of IL-2 (24 h supernatants) and IFN-γ and TNF-α (72 h supernatants) were measured using specific ELISA (eBioscience) according to the manufacturers' recommendations.
5
Cytokine Quantification in Murine Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Levels of IL-5 and IL-13 in MLN cell culture supernatants were measured by specific ELISA (eBioscience, San Diego, CA) as per manufacturer’s instructions. Concentrations of cytokines in BAL fluid were measured with the Bio-plex Pro mouse cytokine 23-plex assay (Biorad, Hercules, CA), with custom made addition of IL-33, and performed according to manufacturer’s instruction. Analysis of the results was performed in Graph Pad Prism. TSLP concentration in BAL fluid and IL-17 concentration in serum were determined with mouse TSLP ELISA read-set-go and mouse IL-17A ELISA read-set-go respectively, both from eBioscience (San Diego, CA), according to manufacturer instructions. Standard curves and analysis of the results were performed with Graph Pad Prism.
6
Immunoglobulin and Cytokine Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sera and peritoneal exudates were analysed for the presence of the various immunoglobulin (Ig) classes (IgM, IgG, IgE and IgA) by ELISA as previously described [8 (link), 18 (link)]. IL-6, IL-10, IL-12/23, IL-21, TNF-α and INF-γ levels were determined in peritoneal exudates using specific ELISA (e-Bioscience). All samples were appropriately diluted before each assay and data were corrected by the dilution factor. Repeated ELISA determinations were submitted to a statistical analysis by using the Mann- Whitney U-test.
7
Extracting Immune Responses from Lung
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood was collected from the iliac vein for serum preparation. BAL fluid was obtained by injecting 3 3 1 mL of PBS containing EDTA into the cannulated trachea. MLN cells (2 3 10 6 cells/mL) were restimulated for 3 days ex vivo with 15 mg/mL HDM extracts in 96 round-bottom plates. Cytokine production was determined in supernatants by using specific ELISA (eBioscience).
8
Biomarker Measurement in EDTA and Heparin Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
CRP, haptoglobin, fibronectin and SAA levels were determined in EDTA and heparinised plasma by means of particle-enhanced immunonephelometry using the BN II System (Siemens Healthcare Diagnostics, Marburg, Germany). Serum IL-6 was detected with a specific ELISA (eBioscience, Vienna, Austria). hsTnT and NT-proBNP measurements were performed in heparin plasma using the Elecsys Systems (Roche Diagnostics, Mannheim, Germany). MR-proANP, MR-proADM, CT-proET-1 and copeptin were measured in EDTA plasma using specific sandwich immunoassays (BRAHMS, Hennigsdorf/Berlin, Germany).
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!