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Dy682

Manufactured by R&D Systems
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Sourced in United States

The DY682 is a laboratory equipment product manufactured by R&D Systems. It is designed to perform a specific function within the research and development laboratory setting. The core function of the DY682 is to assist in the analysis and processing of samples, but no further details about its intended use or capabilities are provided in this factual and unbiased description.

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Lab products found in correlation

Ab100545, by Abcam (1 mentions) Dy871, by R&D Systems (1 mentions) Dy008, by R&D Systems (1 mentions) 8 bromoadenosine 3 5 cyclic monophosphate camp, by Merck Group (1 mentions) Elh pp14, by RayBiotech (1 mentions) Progesterone p4, by Merck Group (1 mentions) β oestradiol e2, by Merck Group (1 mentions)

5 protocols using dy682

1

Quantification of hGH, hPRL, and hIGF1

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Cells were grown to 90% confluence in six-well plates. The medium was then changed to serum-free medium for 48h. Quantification of hGH (ab100526, Abcam) and hPRL (DY682, R&D Systems, Minneapolis, MN, USA) in cell culture conditioned medium and mouse serum hIGF1 (ab100545, Abcam) levels were performed with commercially available ELISA kits as manufactures' instructions.
Kong X., Wu W., Yuan Y., Pandey V., Wu Z., Lu X., Zhang W., Chen Y., Wu M., Zhang M., Li G., Tan S., Qian P., Perry J.K., Lobie P.E, & Zhu T. (2016). Human growth hormone and human prolactin function as autocrine/paracrine promoters of progression of hepatocellular carcinoma. Oncotarget, 7(20), 29465-29479.
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2

Endometrial Stromal Cell Decidualization

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Endometrial co-culture decidualization was assessed by stimulation with 0.5 mM 8-bromoadenosine 3′,5′-cyclic monophosphate (Sigma B5386) and 1 μM medroxyprogesterone 17-acetate (MPA) (Sigma M1629) in the apical and basal medium starting on days 9 and 12 of co-culture, respectively, according to common protocols previously described.39 (link),40 (link) Apical medium (100 μL) was collected on days 5, 7, 9, 12, and 15 of co-culture and stored at −80 °C until analyzed by ELISA for proteins IGFBP-1 (R&D systems DY871) and prolactin (R&D systems DY682), indicative of secretory differentiation. Protocols provided by the manufacturer were adapted to allow ELISAs to be performed in a 384-well plate (ThermoFisher 464718) to minimize medium (sample) volume needed (see Supplementary Information). For primary patient endometrial stromal cell (ESCs) monocultures, cells were incubated with hormone-containing media on days 0, 3, and 6 of culture, and apical conditioned media (100 μL) samples were collected on days 3, 6 and 9 of culture. Reported values are the mean of 3–6 (apical samples) biological replicates (different gels) minus mean concentration of no-cell gel controls.
Cook C.D., Hill A.S., Guo M., Stockdale L., Papps J.P., Isaacson K.B., Lauffenburger D.A, & Griffith L.G. (2017). Local Remodeling of Synthetic Extracellular Matrix Microenvironments by Co-cultured Endometrial Epithelial and Stromal Cells Enables Long-term Dynamic Physiological Function. Integrative biology : quantitative biosciences from nano to macro, 9(4), 271-289.
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3

Quantifying Prolactin in Cell Cultures

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Prolactin ELISA was completed according to manufacturer’s instructions (R&D Systems DY682 and DY008). Briefly, conditioned media from primary human endometrial stromal cells were collected and spun at 590g for 5 minutes at room temperature to pellet cell debris. Prolactin standards, blank, and neat conditioned media were loaded (100 μL) in duplicate. The plate was read at 450 nm on a ClarioStar plate reader (BMG), then analyzed by correcting absorbance values to the blank and performing 4-parameter logistic regression analysis.
Griffiths M.J., Marshall S.A., Cousins F.L., Alesi L.R., Higgins J., Giridharan S., Sarma U.C., Menkhorst E., Zhou W., Care A.S., Donoghue J.F., Holdsworth-Carson S.J., Rogers P.A., Dimitriadis E., Gargett C.E., Robertson S.A., Winship A.L, & Hutt K.J. (2023). Radiotherapy exposure directly damages the uterus and causes pregnancy loss. JCI Insight, 8(6), e163704.
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4

Differentiation of Endometrial Scaffolds

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For the differentiation of both stromal-only and EO-only scaffolds, 10 nM β-oestradiol (E2, Sigma) was added 2 days post-cell seeding. For stromal cells only, at days 4 and 6, medium was replaced with the following conditions: 10 nM E2 + 1 μM progesterone (P4, Sigma) + 1 μM 8-bromoadenosine 3′,5′-cyclic monophosphate (cAMP, Sigma). For EO-only scaffolds, 20 ng ml−1 prolactin (PRL, Peprotech, UK) was added in addition at days 4 and 6 to mimic signals from decidualized stromal cells. Supernatant was collected at days 2, 4, 6 and 8 and frozen at −20°C for storage before downstream analysis.
Enzyme-linked immunosorbent assays (ELISAs) were used to measure protein concentrations of key markers of stromal decidualization (prolactin, a hormone secreted by decidual stromal cells) and epithelial differentiation (glycodelin, one of the principal components of the endometrial glands) in the supernatant. ELISAs for prolactin (DY682, R&Dsystems, USA) and glycodelin (ELH-PP14, Raybiotech, USA) were measured from stromal-only and EO-only scaffolds, respectively. A volume of 100 μl of supernatant was used with each sample measured in duplicate, as per the manufacturer's instructions.
Abbas Y., Brunel L.G., Hollinshead M.S., Fernando R.C., Gardner L., Duncan I., Moffett A., Best S., Turco M.Y., Burton G.J, & Cameron R.E. (2020). Generation of a three-dimensional collagen scaffold-based model of the human endometrium. Interface Focus, 10(2), 20190079.
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5

Quantifying Secreted dPRL by ELISA

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Concentrations of secreted dPRL in cell culture media were measured using commercial ELISA kits according to the manufacturer’s instructions (DY682; R&D systems). Measurements were performed in triplicate.
James D.W., Quintela M., Lucini L., Alkafri N.K., Healey G.D., Younas K., Bunkheila A., Margarit L., Francis L.W., Gonzalez D, & Conlan R.S. (2024). Homeobox regulator Wilms Tumour 1 is displaced by androgen receptor at cis-regulatory elements in the endometrium of PCOS patients. Frontiers in Endocrinology, 15, 1368494.
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