Dang10

Manufactured by R&D Systems

Sourced in United States

The DANG10 is a laboratory instrument designed for the detection and quantification of analytes in biological samples. It utilizes a sensitive and specific detection method to provide accurate and reliable results. The core function of the DANG10 is to facilitate the analysis of target molecules in a controlled and efficient manner, without making claims about its intended use or applications.

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Lab products found in correlation

Dang20, by R&D Systems (4 mentions) Ek1296, by Boster Bio (2 mentions) Proteome profiler mouse xl cytokine array, by R&D Systems (2 mentions) Microplate reader, by Agilent Technologies (2 mentions) Dmp900, by R&D Systems (1 mentions) Dte200, by R&D Systems (1 mentions) Imagequant las 4000, by GE Healthcare (1 mentions) Ab234559, by Abcam (1 mentions) Ab108918, by Abcam (1 mentions) Xt 2000i analyzer, by Sysmex (1 mentions) Qiaamp minelute virus spin kit, by Qiagen (1 mentions) Lightcycler 480, by Roche (1 mentions) Imagequant las 4000 imaging system, by GE Healthcare (1 mentions) Imagequant tl, by GE Healthcare (1 mentions)

8 protocols using dang10

Measuring Angiogenic Biomarkers in Serum

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Peripheral blood samples were collected from all participants at recruitment as part of standard care and serum was processed by the Townsville University Hospital Pathology Department (Pathology Queensland) prior to storing at − 80 °C for later analysis. Serum angpt-1, matrix-metalloproteinase-9 (MMP-9) and Tie-2 concentrations were measured using commercially available ELISAs according to manufacturer’s directions (DANG 10, DMP900 and DTE200, respectively, R&D Systems, Minneapolis, USA). Serum angiopoietin-2 (angpt-2), and vascular endothelial growth factor (VEGF)-A, -C and -D concentrations were measured using the MilliPLEX MAP Human Angiogenesis/Growth Factor Magnetic Bead Panel—Cancer Multiplex Assay kit (HAGP1MAG-12 K, Merck, Australia), using the MagPIX platform, and analyses were conducted by a researcher blinded to patient diagnosis. Samples in which the assessed biomarker fell outside of the detectable limits of the test were excluded from analysis (detailed in Supplement 1). Biomarker analysis was conducted independently of patient care and had no potential to influence clinical outcome.

Moxon J.V., Kraeuter A.K., Phie J., Juliano S., Anderson G., Standley G., Sealey C., White R.P, & Golledge J. (2022). Serum angiopoietin-1 concentration does not distinguish patients with ischaemic stroke from those presenting to hospital with ischaemic stroke mimics. BMC Cardiovascular Disorders, 22, 462.

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Cytokine and ANGPT1 Profiling in Platelets

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Cytokine profiling assays were performed using the Proteome Profiler Mouse XL Cytokine Array (catalo no. ARY028; R&D Systems) according to the manufacturer’s protocol. Briefly, after isolating and activating the platelets of Pf4-Cre⁺Gucy1b1^+/LoxP and Pf4-Cre⁺Gucy1b1^LoxP/LoxP mice by shaking in RPMI as described above, samples were added to the supplied antibody-spotted nitrocellulose membrane and incubated at 4 °C overnight. Captured proteins were detected using a mixture of biotinylated detection antibodies followed by streptavidin-HRP and visualized using chemiluminescent detection reagents. Signal intensities were detected by an ImageQuant LAS 4000 imaging system and analyzed using the appropriate image analysis software (ImageQuant LAS TL, version 8.1; GE Healthcare Life Sciences). The signal intensities of target proteins were normalized to the signal intensities for the reference spots in each sample. The procedure was repeated for a total of six samples per group.
ANGPT1 ELISAs were performed to determine murine (catalog no. EK1296; Boster Biological Technology) and human (catalog no. DANG10; R&D Systems) protein levels according to the manufacturer’s recommendations.

Mauersberger C., Sager H.B., Wobst J., Dang T.A., Lambrecht L., Koplev S., Stroth M., Bettaga N., Schlossmann J., Wunder F., Friebe A., Björkegren J.L., Dietz L., Maas S.L., van der Vorst E.P., Sandner P., Soehnlein O., Schunkert H, & Kessler T. (2022). Loss of soluble guanylyl cyclase in platelets contributes to atherosclerotic plaque formation and vascular inflammation. Nature cardiovascular research, 1(12), 1174-1186.

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Quantifying Angiopoietin Levels in Plasma

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To detect the levels of angiopoietin-1 and angiopoietin-2 in plasma, enzyme-linked immunosorbent assay (ELISA) kits were used and carried out according to the manufacturer’s protocol (DANG10 and DANG20, R&D, MN, USA). Briefly, standards of angiopoietin-1 and angiopoietin-2 were prepared, and 1:5 diluted plasma samples were pipetted into wells with specific monoclonal antibody pre-coated microplate. After washing away unbound solution, enzyme-linked monoclonal antibodies were added to wells. Following a wash to remove any unbound antibody-enzyme solutions, a substrate solution was added to wells and the color developed in proportion to the control or sample concentrations. Finally, a tetramethylbenzidine solution was added to stop the reaction. We detected the intensity of the reaction color at 450 nm wavelength with a microplate reader (BioTek Instruments, VT, USA). All measurements were performed in duplicate.

Wu H.L., Chu Y.H., Tai Y.H., Tsou M.Y., Wu C.H., Lo W.L., Tai S.K., Yeh C.C, & Lu C.C. (2020). Stage-dependent angiopoietin-Tie2 and nitric oxide signaling of erythrocytes in response to surgical trauma in head and neck cancer. World Journal of Surgical Oncology, 18, 209.

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Plasma Biomarkers in Clinical Study

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All routine lab tests were performed at the hospital’s clinical chemistry department. ELISA’s were used to measure the plasma protein concentrations for ANGPT2 (DANG20, R&D Systems, Minneapolis, MN, USA), ANGPT1 (DANG10, R&D Systems, Minneapolis, MN, USA), von Willebrand factor (ab108918, Abcam, Cambridge, UK), and ADAMTS13 (ab234559, Abcam, Cambridge, UK), according to the manufacturer’s instructions. Data for thrombin–antithrombin (TAT) complexes were available for some patients from a previous study [10 (link)].

Hultström M., Fromell K., Larsson A., Persson B., Nilsson B., Quaggin S.E., Betsholtz C., Frithiof R., Lipcsey M, & Jeansson M. (2022). Angiopoietin-2 Inhibition of Thrombomodulin-Mediated Anticoagulation—A Novel Mechanism That May Contribute to Hypercoagulation in Critically Ill COVID-19 Patients. Biomedicines, 10(6), 1333.

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ELISA Quantification of Plasma Biomarkers

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Plasma sPLA-2 Type IIA, Ang1 and Ang2 was measured by ELISA kit (Cayman Chemical: #501380; R&D Systems: #DANG10, #DANG20, respectively), as per manufacturer instructions. Samples were run in duplicate with absorbance read at 450 nm in real time using a spectrophotometer. sPLA-2, Ang1 and Ang concentrations were generated from a seven-point four-parameter logistic standard curve.

Fidanza M., Hibbert J., Acton E., Harbeson D., Schoeman E., Skut P., Woodman T., Eynaud A., Hartnell L., Brook B., Cai B., Lo M., Falsafi R., Hancock R.E., Chiume-Kayuni M., Lufesi N., Popescu C.R., Lavoie P.M., Strunk T., Currie A.J., Kollmann T.R., Amenyogbe N, & Lee A.H. (2024). Angiogenesis-associated pathways play critical roles in neonatal sepsis outcomes. Scientific Reports, 14, 11444.

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Quantifying Angiopoietin Levels in Plasma

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To detect the levels of angiopoietin-1 and angiopoietin-2 in plasma, enzyme-linked immunosorbent assay (ELISA) kits were used and carried out according to the manufacturer's protocol (DANG10 and DANG20, R&D, MN, USA). Brie y, standards of angiopoietin-1 and angiopoietin-2 were prepared, and 1:5 diluted plasma samples were pipetted into wells with speci c monoclonal antibody pre-coated microplate. After washing away unbound solution, enzyme-linked monoclonal antibodies were added to wells. Following a wash to remove any unbound antibody-enzyme solutions, substrate solution was added to wells and color developed in proportion to the control or sample concentrations. Finally, tetramethylbenzidine solution was added to stop the reaction. We detected the intensity of the reaction color at 450 nm wavelength with a microplate reader (BioTek Instruments, VT, USA). All measurements were performed in duplicate.

Wu H., Chu Y., Tai Y., Tsou M., Tsai C., Lo W., Tai S., Yeh C., & Lu C. (2020). Angiopoietin-Tie2 and nitric oxide signaling of erythrocytes in response to surgical trauma in head and neck cancer.

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Biomarker Analysis of COVID-19

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Nasopharyngeal swabs sample handling and analysis were done as described before. 19 (link) In short, extraction of genetic material was performed with the QIAamp MinElute Virus Spin Kit (Qiagen) and detection of viruses was performed with the Respifinder SMART 22 multiplex PCR kit (Pathofinder BV) on a Lightcycler 480 (Hoffmann La Roche), according to the manufacturers' instructions. Blood samples were collected in serum microtainers using venipuncture, capillary collection, or blood collection during the insertion of a venous cannula. Total cell counts were measured routinely with a Sysmex XT 2000i analyzer (Sysmex). To obtain serum, blood was allowed to clot at room temperature and serum was separated by centrifugation at 2300g for 8 min and stored at -80°. Analysis was performed after only one freeze-thaw cycle to prevent stability issues of the biomarkers. Levels of Ang-1 and Ang-2 were measured using ELISA (R&D Systems DANG10 and DANG20 Quantikine ELISAs). Levels of sE-selectin, sP-selectin, soluble vascular cell adhesion molecule (VCAM-1), and soluble intercellular adhesion molecule (ICAM-1) were measured using Luminex (Human Adhesion 6-plex; Thermo Fisher Scientific). All assays were performed according to the manufacturers' instructions. Ang-2/Ang-1 ratios and NLR were calculated.

Juliana A., Plötz F.B., Achten N., Bultman A., Jongman R.M., van Meurs M., Wilschut J.C, & Zonneveld R. (2021). Requirement of respiratory support in acute bronchiolitis in infants is linked to endothelial and neutrophil activation. Pediatric pulmonology, 56(12).

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Proteome Profiling and Angiopoietin-1 ELISA

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Proteome profiling assays were performed using the Proteome Profiler Mouse XL Cytokine Array (ARY028, R&D Systems) according to the manufacturer's protocol. Briefly, after isolating and activating platelets of Pf4-Cre + Gucy1b1 +/flox mice and Pf4-Cre + Gucy1b1 flox/flox mice by shaking in RPMI as described above, samples were added to the supplied antibodyspotted nitrocellulose membrane and incubated at 4°C overnight. Captured proteins were detected using a mixture of biotinylated detection antibodies followed by streptavidinhorseradish peroxidase and visualized using chemiluminescent detection reagents. Signal intensities were detected by an ImageQuant LAS 4000 imaging system (GE Healthcare Life Sciences, Freiburg, Germany) and analyzed using the appropriate image analysis software (ImageQuant TL; GE Healthcare Life Sciences). Signal intensities of target proteins were normalized to signal intensities for reference spots in each sample. The procedure was repeated for a total of 6 samples per group.
Angiopoietin-1 enzyme-linked immunosorbent assays (ELISA) were performed to determine murine (EK1296, Boster Biological Technology, Pleasanton, CA) and human (DANG10, R&D systems) protein levels according to the manufacturer's recommendations.

Mauersberger C., Sager H.B., Wobst J., Dang T.A., Lambrecht L., Koplev S., Stroth M., Bettaga N., Schlossmann J., Wunder F., Friebe A., Björkegren J., Dietz L., Sandner P., Soehnlein O., Schunkert H., & Kessler T. (2021). Loss of soluble guanylyl cyclase in platelets contributes to atherosclerotic plaque formation and vascular inflammation.

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