Histopaque-1077 and −1119, RPMI 1640, Luria-Bertani (LB) broth, bovine serum albumin (BSA), paraformaldehyde, nalidixic acid and kanamycin were procured from Sigma (St. Louis, MO). DNase I (dornase alfa, Pulmozyme®) was kindly gifted from Genentech. Duoset enzyme-linked immunosorbent assay (ELISA) kits for mouse Lipocalin2 (Lcn2), keratinocyte-derived chemokine (KC), serum amyloid A (SAA), neutrophil elastase (NE), myeloperoxidase (MPO), interleukin (IL)-22 and IL-17A were obtained from R&D Systems (Minneapolis, MN). SYBR Green mix and the qScript complementary DNA (cDNA) synthesis kit were procured from Quanta BioSciences (Gaithersburg, MD). Ly6G antibody was purchased from Abcam (Cambridge, MA). All other fine chemicals used in the present study were reagent grade and procured from Sigma. Alcian Blue (pH 2.5) Stain Kit was obtained from Vector Laboratories (Burlingame, CA). Mouse Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec (Auburn, CA). Cl-amidine (hydrochloride) was obtained from Cayman chemicals (Ann Arbor, MI).
Duoset enzyme linked immunosorbent assay
Manufactured by R&D Systems
Sourced in United States, United Kingdom, Niger
The DuoSet enzyme-linked immunosorbent assay (ELISA) is a laboratory product designed for the quantitative measurement of specific proteins or other analytes in biological samples. It provides a standardized and reliable platform for performing ELISA assays.
Automatically generated - may contain errors
Lab products found in correlation
Bovine serum albumin (bsa), by Merck Group (3 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Histopaque 1077 and 1119, by Merck Group (2 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Kanamycin, by Merck Group (2 mentions)
Luria bertani broth, by Merck Group (2 mentions)
Nalidixic acid, by Merck Group (2 mentions)
Interleukin il 22, by R&D Systems (2 mentions)
Sybr green mix, by Quanta Biosciences (2 mentions)
Qscript complementary dna cdna synthesis kit, by Quanta Biosciences (2 mentions)
Ly6g antibody, by Abcam (2 mentions)
Alcian blue ph 2.5 stain kit, by Vector Laboratories (2 mentions)
Cl amidine hydrochloride, by Cayman Chemical (2 mentions)
Neutrophil elastase, by R&D Systems (2 mentions)
Mouse lamina propria dissociation kit, by Miltenyi Biotec (2 mentions)
Il 17a, by R&D Systems (2 mentions)
Serum amyloid a, by R&D Systems (2 mentions)
2030 arvo x4, by PerkinElmer (1 mentions)
Elisa development system, by R&D Systems (1 mentions)
Architect immunoassay, by Abbott (1 mentions)
20 protocols using duoset enzyme linked immunosorbent assay
1
Extraction and Quantification of Murine Immune Factors
2
Quantification of Inflammatory Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of MPO, MMPs and the cytokines IL-1β, IL-8, and tumor necrosis factor (TNF)-α in cell culture supernatants were measured using the DuoSet enzyme-linked immunosorbent assay (ELISA) development system (R&D Systems) and ELISA MAX Standard Set Human TNF-α (Cat# 430201; BioLegend, San Diego, CA, USA) according to manufacturer' instructions. The absorbance at 450 nm was measured on a microplate reader (2030 ARVO X4; PerkinElmer Japan; Tokyo).
3
Comprehensive Biomarker Profiling in Plasma and Urine
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Both urine and plasma samples were used to measure endogenous biomarkers (see Figure 1 for details and abbreviations for each analyte and biomarker). We measured inflammatory markers in plasma samples at the University of Michigan Cancer Center Immunology Core, including four cytokines using the Milliplex Multiplex Assay Simultaneous High Sensitivity Human Cytokine Magnetic Bead Panel (EMD Millipore Corp.) and C-reactive protein using a DuoSet enzyme-linked immunosorbent assay (R&D Systems). We quantified plasma concentrations of the angiogenic biomarkers PGF and sFlt-1 using the ARCHITECT immunoassay (Abbott Laboratories). In addition, we measured a panel of 53 eicosanoids in plasma using a 6490 triple quadrupole mass spectrometer (Agilent). Three unique protein damage markers NY, DY, and CY) were measured in plasma samples using ESI-MS/MS. Finally, two oxidative stress markers were measured in urine samples at Cayman Chemical: 8-IP, which was quantified via affinity column chromatography and enzyme immunoassay, and 8-OHdG, which was quantified with direct dilution and enzyme immunoassay. Endogenous biomarkers that were below the LOD were imputed with the LOD value divided by the square root of 2. More extensive details on analysis and measurement of endogenous biomarkers have been previously described (Aung et al. 2019b (link); Ferguson et al. 2017 (link)).
4
Quantifying cytokine release from M. tuberculosis-infected human macrophages
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An actively growing M. tuberculosis–lux culture (OD, 0.3–0.6) was pelleted at 3500g for 5 minutes, washed twice, and resuspended in Sauton’s medium. An appropriate volume of this inoculum was then applied to differentiated human MDMs using the OD-to-CFU/mL conversion factor above. For these experiments, monolayers were not washed, and supernatants were simply harvested after 24 hours and filtered using a MultiScreenHTS 0.22-μm plate manifold (Millipore) before removal from a biosafety level 3 facility. Cytokine concentrations were assayed by DuoSet enzyme-linked immunosorbent assay (R&D Systems) using 3, 3′, 5, 5′ - tetramethylbenzidine (TMB) peroxidase substrate (SeraCare) and read on a SpectraMax-Plus plate reader (Molecular Devices).
5
Cytokine Measurement by ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IL-1β and TNF-α in conditioned media were measured using the antibodies and reference standards contained in R&D Systems DuoSet enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s protocol.
6
Quantifying Mouse IL-1β and CXCL1 Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse IL-1β and CXCL1 (KC) were measured using DuoSet enzyme-linked immunosorbent assay (ELISA; R&D Systems, Minneapolis, MN).
7
Quantifying Human Cytokine Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human IL-6 and CXCL8/IL-8 DuoSet enzyme linked immunosorbent assay (R and D Systems, Abingdon, United Kingdom) were used in combination with the appropriate DuoSet Ancillary Reagent Kit (R and D Systems), according to manufacturer’s instructions, to detect cytokine protein levels in a sample. Briefly, the supernatant samples were diluted 1:20. Optical density at 450 nm with background correction at 540 nm was determined immediately after the addition of the stop solution. Cytokine concentration was determined using 4-parameter fit standard. Data was corrected for the number of cells using viability data.
8
Quantifying ACE1 and ACE2 in Serum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen (− 20 °C) serum samples of the hospitalized participants were thawed and the concentrations of ACE1 and ACE2 were measured according to manufacturer’s instructions, using the R&D-Systems duoset enzyme-linked immunosorbent assay (ELISA).
9
Extraction and Quantification of Murine Immune Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histopaque-1077 and −1119, RPMI 1640, Luria-Bertani (LB) broth, bovine serum albumin (BSA), paraformaldehyde, nalidixic acid and kanamycin were procured from Sigma (St. Louis, MO). DNase I (dornase alfa, Pulmozyme®) was kindly gifted from Genentech. Duoset enzyme-linked immunosorbent assay (ELISA) kits for mouse Lipocalin2 (Lcn2), keratinocyte-derived chemokine (KC), serum amyloid A (SAA), neutrophil elastase (NE), myeloperoxidase (MPO), interleukin (IL)-22 and IL-17A were obtained from R&D Systems (Minneapolis, MN). SYBR Green mix and the qScript complementary DNA (cDNA) synthesis kit were procured from Quanta BioSciences (Gaithersburg, MD). Ly6G antibody was purchased from Abcam (Cambridge, MA). All other fine chemicals used in the present study were reagent grade and procured from Sigma. Alcian Blue (pH 2.5) Stain Kit was obtained from Vector Laboratories (Burlingame, CA). Mouse Lamina Propria Dissociation Kit was purchased from Miltenyi Biotec (Auburn, CA). Cl-amidine (hydrochloride) was obtained from Cayman chemicals (Ann Arbor, MI).
10
TCR Expression and Activation Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Missing sequence parts of leader, variable and constant regions of selected TCRs were completed with data downloaded from the international ImMunoGeneTics information system. Reconstructed TCR sequences were synthesized (Thermo Fisher Scientific, Waltham, MA, USA) and expressed in 58a À b À cell lines as previously described [32, 33] . The 58a À b À cells expressed human CD8ab chains and green fluorescent protein (GFP) under the control of the nuclear factor of activated T-cell promoter [33] , thus indicating T-cell activation by GFP expression. TCR expression was confirmed by mouse CD3 staining and detection by flow cytometry. As positive control for TCR activation, TCR-recombinant cell lines were stimulated with plate-bound anti-mouse CD3 for 16 h at 37°C and 5% CO 2 . GFP expression was measured with flow cytometry and IL-2 production was detected in cell culture supernatants using the DuoSet enzyme-linked immunosorbent assay (ELISA) ancillary reagent kit 2 (R&D Systems, Minneapolis, MN, USA).
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!