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Dtga00

Manufactured by R&D Systems
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The DTGA00 is a thermal analysis instrument designed to measure the weight change of a sample as a function of temperature and time. It provides quantitative information about physical and chemical changes that occur in materials during heating or cooling.

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Lab products found in correlation

Ab100531, by Abcam (1 mentions) Dar00, by R&D Systems (1 mentions) Deg00, by R&D Systems (1 mentions) Mm200, by R&D Systems (1 mentions) Mkc00b, by R&D Systems (1 mentions)

4 protocols using dtga00

1

Quantification of AREG, TGF-α, EGF, and HB-EGF Secretion

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The concentration of secreted AREG protein in cultured media was determined using the AREG Quantikine ELISA kit (R&D Systems-DAR00). The concentrations of TGF-α (R&D Systems-DTGA00), EGF (R&D Systems-DEG00) and HB-EGF (Abcam-ab100531) were also measured in the same conditioned media using specific Quantikine ELISA kits. In brief, 2×104 cells/well were seeded in the 48-well plates and cultured overnight. Cells were then washed with 1×phosphate-buffered saline (PBS) once and incubated in 0.5 ml of complete medium for 24 h. The media were collected and centrifuged and supernatant was collected and stored at −80°C prior to analysis. Cells were trypsinized and counted. ELISA assays were performed according to the manufacturer's protocols. Samples were run in triplicate, and secreted AREG was normalized against standard curves and presented as pg per 1×104 cells.
Wen Y., Li H., Zeng Y., Wen W., Pendleton K.P., Lui V.W., Egloff A.M, & Grandis J.R. (2016). MAPK1E322K mutation increases head and neck squamous cell carcinoma sensitivity to erlotinib through enhanced secretion of amphiregulin. Oncotarget, 7(17), 23300-23311.
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2

TGF-α and Betacellulin Shedding Assay

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A549 cells were cultured in T175 flasks in F12K media supplemented by 10% FBS. For assay, cells were passaged and seeded at 1.3 × 104 in 24-well plates in serum-free F12K media. All test and control inhibitors were added and incubated for 1 h, after which PMA was added at 500 ng/mL final concentration to stimulate TGFα shedding. In the case of betacellulin, PMA was not added because betacellulin is cleaved constitutively. All leads were tested at 40 μM, and marimastat was tested at 10 μM. A549 cells were incubated with PMA for 1 h, after which supernatant was collected and concentrated using Amicons. TGFα and betacellulin were detected in supernatant by ELISA (R&D Systems catalogue nos. DTGA00 and DY261, respectively).
Knapinska A.M., Dreymuller D., Ludwig A., Smith L., Golubkov V., Sohail A., Fridman R., Giulianotti M., LaVoi T.M., Houghten R.A., Fields G.B, & Minond D. (2015). SAR Studies of Exosite-Binding Substrate-Selective Inhibitors of A Disintegrin And Metalloprotease 17 (ADAM17) and Application as Selective in Vitro Probes. Journal of medicinal chemistry, 58(15), 5808-5824.
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3

ELISA Analysis of Secreted Growth Factors

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HDMEC and tumor cells were cultured with serum free medium (EBM and DMEM, respectively) for 24 hours. The supernatant was collected and ELISA for EGF (Cat. DEG00, R&D Systems), TGF-alpha (Cat. DTGA00, R&D Systems) and Amphiregulin (Cat. DAR00, R&D Systems) was performed.
Zhang Z., Dong Z., Lauxen I.S., Filho M.S, & Nör J.E. (2014). Endothelial cell-secreted EGF induces epithelial to mesenchymal transition and endows head and neck cancer cells with stem-like phenotype. Cancer research, 74(10), 2869-2881.
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4

BALF Cytokine Quantification by ELISA

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The supernatant fraction of the BALF was thawed on ice and used in enzyme-linked immunosorbant assays (ELISA). The mouse homologs of human interleukin (IL)-8, Keratinocyte-Derived Cytokine (KC) and Macrophage Inflammatory Protein (MIP)-2, were detected using ELISA (R&D Systems, Product Number MKC00B and MM200, respectively). TGF-α was also analyzed similarly (R&D Systems, Product number DTGA00). BALF cytokine concentrations were determined by comparison to standard curves for each cytokine provided by the supplier.
Chang S., Linderholm A, & Harper R. (2015). DUOX-Mediated Signaling Is Not Required for LPS-Induced Neutrophilic Response in the Airways. PLoS ONE, 10(7), e0131810.
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