Dm3a00

Manufactured by R&D Systems

Sourced in United States

The DM3A00 is a laboratory equipment designed for measurement and analysis. It serves as a tool for collecting and processing data related to various scientific experiments and research activities.

Automatically generated - may contain errors

Lab products found in correlation

Ab222868, by Abcam (1 mentions) Ab193716, by Abcam (1 mentions) Dip100, by R&D Systems (1 mentions) Dgr00, by R&D Systems (1 mentions) Ab184862, by Abcam (1 mentions) Ascent software version 2, by Thermo Fisher Scientific (1 mentions) Det100, by R&D Systems (1 mentions) Dmd00, by R&D Systems (1 mentions) Dcp00, by R&D Systems (1 mentions) Dcc700, by R&D Systems (1 mentions)

6 protocols using dm3a00

ELISA Quantification of CCL20 and CXCL8

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Basal media were used for ELISA quantification of human CCL20 and CXCL8 following the instructions of the commercial kit (DM3A00 and D8000C, respectively, R&D systems).

Rahmawati S.F., Vos R., Bos I.S., Kerstjens H.A., Kistemaker L.E, & Gosens R. (2022). Function-specific IL-17A and dexamethasone interactions in primary human airway epithelial cells. Scientific Reports, 12, 11110.

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Multiplexed Cytokine and ELISA Assays

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All samples were stored at −80 °C before they were diluted 1:2 for Cytokine analysis using Human Magnetic Luminex Assay (PN LXSAHM-19; R&D Systems) with the manufacturer’s set protocol. Data was collected on a Luminex 200 (Luminex) and analyzed on Milliplex analyst software (Vigene tech). In addition, after 1X thaw, samples were tested with individual Quantitative ELISA Kit per manufacturer’s protocol as follows: Human Thrombospondin 1:20(PN ab193716; Abcam); SLP-I 1:20(PN DP100; R&D Systems); Annexin A1 1:20(PN ab222868; Abcam); IL-8/CXCL8 1:20(PN D8000C; R&D Systems); CCL20/MIP-3 alpha 1:20(PN DM3A00; R&D Systems); CXCL10/IP-10 1:50(PN DIP100; R&D Systems); Human EN-RAGE 1:100(PN D41052-05; R&D Systems); Human S100A8/S100A9 Heterodimer 1:1000(PN D48226-05; R&D Systems). After 2× thaw samples were diluted for another set of individual Quantitative ELISA Kits as follows; Human FOLR3 1:20(PN DY5319; R&D Systems); Human CCL3L1/LD78 Beta 1:20(PN NBP2-7065; Novus); Human CCL4L1/LAG-1 1:20(PN NBP2-75067; Novus). All ELISAs were read on ELISA Reader (Molecular Probes) using dual wavelengths of 450/540 and data was analyzed using SOFTMAX Pro software 7.1.4.

Flerlage T., Crawford J.C., Allen E.K., Severns D., Tan S., Surman S., Ridout G., Novak T., Randolph A., West A.N, & Thomas P.G. (2023). Single cell transcriptomics identifies distinct profiles in pediatric acute respiratory distress syndrome. Nature Communications, 14, 3870.

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Measuring Chemokines and Cholesterol

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The concentrations of the specific chemokines and cholesterol in the supernatant of cell culture or mouse serum were measured using ELISAs according to the manufacturer’s instructions. The ELISA kits were human CXCL1 (DGR00, R&D Systems), human CXCL2 (ab184862, Abcam), human CXCL8 (D8000C, R&D Systems), human CCL20 (DM3A00, R&D Systems), and human cholesterol (MBS729689, Mybiosource).

Han B., Alonso-Valenteen F., Wang Z., Deng N., Lee T.Y., Gao B., Zhang Y., Xu Y., Zhang X., Billet S., Fan X., Shiao S., Bhowmick N., Medina-Kauwe L., Giuliano A, & Cui X. (2021). A chemokine regulatory loop induces cholesterol synthesis in lung-colonizing triple-negative breast cancer cells to fuel metastatic growth. Molecular Therapy, 30(2), 672-687.

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Cytokine and Mediator Quantification

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Quantification of macrophage-derived chemokine (MDC or CCL22), CCL20, ET-1, and IL-10 was performed using ELISA kits from R&D Systems (DMD00, DM3A00, DET100, and D1000 B, respectively, Minneapolis, MN, USA). Quantification of TXA 2 , IL-33, and calcitonin was performed using ELISA kits from Biomatik (EKU07649, Kitchener, Ontario, Canada), abcam (ab108918, Cambridge, UK), Thermo Scientific (EHIL33, Waltham, MA, USA), and Cusabio (CSB-E05131h, Houston, TX, USA), respectively. All ELISAs were performed according to the manufacturer's instructions. The optical densities (O.D.) were read at 450 nm with a wavelength correction at 540 nm in a Multiscan Ascent V1.24 with Ascent Software version 2.6 (Thermo Scientific, Waltham, MA, USA). Data values were expressed as pg/mL deduced from the standard curve after subtracting the blanks, using a 4parameter logistic algorithm.

Vihlborg P., Lundberg O., Pettersson-Pablo P., Johansson N., Bryngelsson I.L., Stjernbrandt A, & Graff P. (2024). Blood biomarkers for occupational hand-arm vibration exposure. Toxicology and industrial health, 40(8).

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CCL20 Quantification by ELISA

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CCL20 was measured by using specific ELISA kits with commercially available paired antibodies (R&D Systems, DM3A00). Specific methods were carried out in accordance with the manufacturer’s instructions. The sensitivity of the CCL20 ELISA was 0.87 pg/mL.

Lu G., Zhang X., Shen L., Qiao Q., Li Y., Sun J, & Zhang J. (2017). CCL20 secreted from IgA1-stimulated human mesangial cells recruits inflammatory Th17 cells in IgA nephropathy. PLoS ONE, 12(5), e0178352.

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Quantifying Chemokine Secretion in HUVECs

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For detection of chemokine secretion, HUVECs were left unstimulated or stimulated with TNF-α and/or IFN-γ for 18 h in culture conditions as described above. Medium was collected, and cells were lysed in medium containing 0.1% Triton X-100. Concentrations of chemokines were determined using ELISA kits from R&D Systems (CCL2; DCP00, CCL7; DCC700, CCL8; DY281, CCL20; DM3A00) according to the manufacturer’s instructions. Final values were averages of measurements performed in duplicate or triplicate. Levels of chemokines in the conditioned medium from cultured cells and the cell lysate were determined by reference to a standard curve produced using recombinant protein. To measure apically and basally secreted chemokines, HUVEC monolayers were grown overnight in culture conditions as described above but on fibronectin coated transwells (3415; Corning) with a 3 μm pore size. Monolayers were washed gently and activated with TNF-α and IFN-γ and conditioned medium was collected from upper and lower wells after 4 h. The upper well samples were diluted with medium to equalize sample volumes and chemokine concentrations were measured by ELISA.

Parween F., Singh S.P., Zhang H.H., Kathuria N., Otaizo-Carrasquero F.A., Shamsaddini A., Gardina P.J., Ganesan S., Kabat J., Lorenzi H.A., Myers T.G, & Farber J.M. (2023). Chemokine positioning determines mutually exclusive roles for their receptors in extravasation of pathogenic human T cells. bioRxiv.

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