Commercially available ELISA kits were used to measure the concentrations or activity of IFNβ (DIFNB0 or 42400–1, R&D Systems), F3 (DCF300, R&D Systems; ab214091, Abcam), fibrin (MBS265263 or MBS706338, MyBioSource), D-dimer (ab196269, Abcam; MBS723281, MyBioSource), TNF (DTA00D, R&D Systems), IL1A (DLA50, R&D Systems), IL1B (DLB50, R&D Systems), IL6 (D6050, R&D Systems), and HMGB1 (ST51011, IBL International) in the indicated samples. Measurement of GPT/ALT and BUN in the plasma was performed using an IDEXX Catalyst Dx Chemistry Analyzer. PT, APTT, and fibrinogen were measured in an automated coagulometer (Sysmex CA-7000). Platelet count was measured using an IDEXX ProCyte Dx Hematology Analyzer.
Dta00d
Manufactured by R&D Systems
Sourced in United States
The DTA00D is a laboratory instrument designed for differential thermal analysis (DTA). It measures the temperature difference between a sample and an inert reference material as a function of time or temperature under controlled atmospheric conditions.
Automatically generated - may contain errors
Lab products found in correlation
D6050, by R&D Systems (16 mentions)
Dlb50, by R&D Systems (12 mentions)
Mlb00c, by R&D Systems (2 mentions)
Db100b, by R&D Systems (2 mentions)
Dif50c, by R&D Systems (2 mentions)
D2050, by R&D Systems (2 mentions)
Mta00b, by R&D Systems (2 mentions)
Mkc00b, by R&D Systems (2 mentions)
Ab214091, by Abcam (1 mentions)
Ab196269, by Abcam (1 mentions)
Difnb0, by R&D Systems (1 mentions)
Dcf300, by R&D Systems (1 mentions)
Mbs723281, by MyBioSource (1 mentions)
Dla50, by R&D Systems (1 mentions)
Catalyst dx chemistry analyzer, by IDEXX (1 mentions)
Procyte dx hematology analyzer, by IDEXX (1 mentions)
Ca 7000, by Sysmex (1 mentions)
Dip100, by R&D Systems (1 mentions)
Ifn α, by R&D Systems (1 mentions)
Ifn β, by R&D Systems (1 mentions)
33 protocols using dta00d
1
Biomarker Quantification in Plasma
2
Cytokine and cGAMP Induction by CX-5461
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HT-29, DLD-1, and CT-26 cells were seeded in a 24-well plate in the absence or presence of 5 µM CX-5461. After incubation at 37°C for 0, 3, 6, 12, 18, and 24 h, the culture media were collected, and the concentrations of type I IFNs were determined using the following enzyme-linked immunosorbent assay (ELISA) kits: IFN-α (human), R&D # 41100-1; IFN-α (mouse), R&D # 42120-1; IFN-β (human), R&D # DIFNB0; IFN-β (mouse), R&D # MIFNB0. To analyze other cytokines, cells were seeded in the absence or presence of 1 and 5 µM CX-5461 for 48 h. Next, the culture media were collected, and the concentrations of TNF-α, IL-6, and CXCL10 were measured using the following ELISA kits: R&D #DTA00D, R&D #D6050, and R&D #DIP100, respectively. To measure cGAMP, the cells were collected and resuspended in RIPA buffer. After centrifugation, the cGAMP levels in the supernatant were determined using a 2′,3′-Cyclic GAMP ELISA Kit (ArborAssay #K067-H1). The results were calculated using an online tool provided from MyAssays (www.myassays.com/arbor-assays-detectx-2-3-cyclic-gamp-eia-kit.assay ).
3
Evaluating Anti-Inflammatory Effects of Dapagliflozin in HK-2 Cells
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HK-2 cells were cultured in K-SFM serum-free medium in six-well plates. When the cells were 70–80% confluent, cells were divided into 3 groups: control (Cont), HG (30 mM), and HG (30 mM) with dapagliflozin (DAPA, 20 μM). After 48 h, supernatants of cultured medium were collected. Levels of pro-inflammatory cytokines in the cultured medium were detected by using human ELISA kits targeting IL-1β (DLB50, R&D, Minneapolis, MN, USA), IL-6 (D6050, R&D, Minneapolis, MN, USA), and TNFα (DTA00D, R&D, Minneapolis, MN, USA), according to the manufacturer’s instructions. The optical densities (O.D.) of samples were measured and corrected by subtracting the readings at 570 nm from the readings at 450 nm using an Emax Plus Microplate Reader (Molecular Devices, San Jose, CA, USA). The concentrations of pro-inflammatory cytokines in cultured medium were calculated by the standard curve and the O.D. of samples.
4
Quantification of Inflammatory Markers
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Quantification of IL-6, IL-1β, TNF-α and CCL2s in serum samples was performed by ELISA, employing kits from R&D Systems (D6050, DLB50, DTA00D and DCP00, respectively). Highly purified recombinant proteins were used to generate six-point standard curves (concentration range between 100 pg/ml and 3.13 pg/ml for IL-6; between 125 pg/ml and 3.9 pg/ml for IL-1β; between 1000 pg/ml and 15.6 pg/ml for TNF-α and between 1000 pg/ml and 31.3 pg/ml for CCL2). When specified in the manufacturer's protocol, serum samples were diluted 2-fold with the appropriate Calibrator Diluent. For each ELISA, 200 μl of sample or calibrator were loaded in each well of the provided 96-well plate. The optical density in each well was determined at 450 nm, using a SpectraMax Plus spectrophotometer (Molecular Devices), with wavelength correction set at 570 nm. A standard curve was generated for each protein using a four-parameter logistics (4-PL) curve-fit and, for each sample, the obtained concentrations of each cytokine/chemokine were multiplied by the appropriate dilution factor. Results are presented in pg/ml of serum.
5
Myocardial Cytokine Quantification
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Interleukin (IL)‐6, IL‐1β, and tumor necrosis factor (TNF)‐α levels in the homogenized myocardial tissue and cell supernatant were measured with ELISA kits (MLB00C, D6050, and DTA00D; R&D Systems) according to the manufacturer's instructions. Briefly, the tissue homogenate was processed by centrifugation at 1000×g for 10 min at 4°C. The supernatants were then cultured with the ELISA reagent. Finally, the optical density (OD) was measured using a microplate reader (Multiskan MK33; Thermolab Systems).
6
Amisulpride Modulates Chemokine Production in Synovial Fibroblasts
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SFs at passage 3 isolated from 8-week-old WT or hTNFtg ankle joints were starved overnight and were treated with different amisulpride concentrations. WT SFs were also stimulated with 10 ng/mL hTNF (Peprotech) or hTNF preincubated with amisulpride for 30 minutes. At 48 hours after, the cell culture supernatants were analyzed for the detection of CCL5 (DY478, DuoSet), CCL20 (DY760, DuoSet), hTNF (R&D Systems, DTA00D), or other chemokines (Mouse Legendplex Chemokine Panel, BioLegend, 740007) according to the manufacturer’s instructions. The toxicity of amisulpride was assessed using the crystal violet assay on SFs, as described elsewhere (79 (link)).
7
Preeclampsia and OSA Biomarker Evaluation
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The patient’s whole blood was collected at the time of enrollment. Blood samples were centrifuged, and separated serum were stored at −80ºC until analysis of enzyme-linked immunosorbent assay (ELISA). We measured serum markers related to preeclampsia,22 (link) such as serum soluble fms-like tyrosine kinase-1 (sFlt-1; MBS262972-96T, MyBiosource, San Diego, CA, USA), endoglin (DNDG00, R&D Systems, Minneapolis, MN, USA), and placental growth factor (PlGF; LS-F1231, LSBio, Seattle, WA, USA). Biomarkers of OSA were evaluated, including interleukin (IL)-6 (D6050, R&D Systems), myeloid-related protein (MRP)8/14 (439707, Biolegend, San Diego, CA, USA), and tumor necrosis factor (TNF)-α (DTA00D, R&D Systems).23 (link)
8
Quantification of Immune Mediators
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Culture supernatants were harvested and centrifuged to remove cellular debris, and aliquots were stored at –80° C until assayed. The concentrations of IL-10, Arg1, iNOS, and TNF-α were determined using corresponding human ELISA kits (IL-10: D1000B, R&D Systems; Arg1: P05089, Cusabio, Houston, TX, USA; iNOS: P35228, Cusabio; TNF-α: DTA00D, R&D Systems) following the methods described previously [62 (link)].
9
Serum Biomarker Profiling in Preeclampsia
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Serum samples were collected from patients with sPE or a gestational age-matched normal pregnancy to detect the secreted IL-32 level using an enzyme-linked immunosorbent assay (ELISA) kit purchased from Sino Biological (IL-32 ELISA Kit, Human; Cat: KIT11064). The levels of IL-1β (DLB50, R&D Systems), TNFα (DTA00D, R&D Systems), sICAM-1 (ELH-ICAM1, RayBiotech, Peachtree Corners, GA, USA) and VCAM-1 (ELH-VCAM1, RayBiotech) in culture medium were measured with ELISA kits and conducted according to the standard procedure.
10
Quantification of TNF-α and IL-1β levels
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TNF-α and IL-1β secretion in the culture medium was evaluated by an ELISA commercial kit (DTA00D, R&D Systems, Minneapolis, MN, USA, and 900-K95, Peprotech, Minneapolis, MN, USA, respectively). Both kits had a detection limit of 15 pg/mL. Cytokine levels were normalized per gram of wet tissue.
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