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Dy275

Manufactured by R&D Systems
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The DY275 is a laboratory instrument designed for performing immunohistochemistry (IHC) and in situ hybridization (ISH) assays. It provides automated control of slide processing steps, including deparaffinization, rehydration, antigen retrieval, and reagent application. The DY275 is capable of handling a variety of sample types and can accommodate multiple slide formats.

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Lab products found in correlation

Dy215, by R&D Systems (2 mentions) Dy271, by R&D Systems (2 mentions) Zymosan tlrl zyn, by InvivoGen (2 mentions) Zymosan, by InvivoGen (2 mentions) Dy208, by R&D Systems (2 mentions) Curdlan, by Merck Group (2 mentions) Ary005b, by R&D Systems (1 mentions) 96 well plate, by Corning (1 mentions) Mccoy s 5a modified medium, by Thermo Fisher Scientific (1 mentions) Dy453, by R&D Systems (1 mentions) Multiplex cytokine immunoassay panel, by Mesoscale (1 mentions)

6 protocols using dy275

1

Cytokine Profiling of Cell Lines

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Secreted cytokine levels, normalized to the total cell number, were measured in the supernatant of cell lines after 4 days growth, using the human cytokine array (R&D Systems, ARY005B) and human quantitative IL-1α and CXCL1 ELISA kits (R&D Systems, DY200 and DY275) according to the manufacturer’s instructions.
Enjalbert F., Dewan P., Caley M.P., Jones E.M., Morse M.A., Kelsell D.P., Enright A.J, & O’Toole E.A. (2020). 3D model of harlequin ichthyosis reveals inflammatory therapeutic targets. The Journal of Clinical Investigation, 130(9), 4798-4810.
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2

Inhibition of IL-17A-Induced CXCL1 Secretion

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HT-29 cells (#HTB-38; ATCC) were plated in a 96 well plate (#3596; Costar) at 20,000 cells per well in McCoy’s 5A (Modified) medium (#16600–082; Gibco) supplemented with 10% FBS. Cells were treated with 60 ng/mL (1,875 pM) human IL-17A (#317-ILB; R&D Systems) in the presence of peptide at the indicated concentrations. A dose range of 0.05 nM to 50,000 nM was evaluated. After ~48 hours, CXCL1/GROα in the culture media was measured using a commercial ELISA kit (#DY275; R&D Systems). Medium alone treatments were included in every experiment to assess the basal levels of CXCL1/GROα. Percent inhibition was calculated by subtracting the mean IL-17A alone CXCL1/GROα values and then dividing by the IL-17A alone value (subtracted from the basal CXCL1/GROα level). So the differences observed at the bottom end of the curve are truly peptide-dependent.
Ting J.P., Tung F., Antonysamy S., Wasserman S., Jones S.B., Zhang F.F., Espada A., Broughton H., Chalmers M.J., Woodman M.E., Bina H.A., Dodge J.A., Benach J., Zhang A., Groshong C., Manglicmot D., Russell M, & Afshar S. (2018). Utilization of peptide phage display to investigate hotspots on IL-17A and what it means for drug discovery. PLoS ONE, 13(1), e0190850.
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3

Cytokine-Mediated CXCL1 Induction Assay

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For inhibitor screen, human BJ fibroblast cells (ATCC CRL2522) (American Type Culture Collection, VA) were used. For mouse cell based assay MLE-12 mouse epithelial cells (ATCC CRL2110) were used. Both cell lines were maintained in ATCC recommended media. Cells were seeded at 5 × 103 cells/well into 96-well flat-bottom microtiter plates in which peptides that had been pre-diluted with cytokines (1 ng/mL for human IL-17A or 15 ng/mL for mouse IL-17A) in culture medium. Cells were incubated at 37 °C for 16–24 hrs, and supernatants were collected and analyzed by ELISA for either human CXCL1/GRO-α (R&D Systems DY275) or mouse CXCL1/KC (R&D Systems DY453).
Liu S., Desharnais J., Sahasrabudhe P.V., Jin P., Li W., Oates B.D., Shanker S., Banker M.E., Chrunyk B.A., Song X., Feng X., Griffor M., Jimenez J., Chen G., Tumelty D., Bhat A., Bradshaw C.W., Woodnutt G., Lappe R.W., Thorarensen A., Qiu X., Withka J.M, & Wood L.D. (2016). Inhibiting complex IL-17A and IL-17RA interactions with a linear peptide. Scientific Reports, 6, 26071.
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4

Senescence-Associated Secretome Analysis

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A Meso Scale Discovery multiplex cytokine immunoassay panel was used to quantitate secreted components of the senescence associated secretory phenotype (SASP), including IL6 and CXCL1 according to manufacturer’s instructions (Mesoscale Diagnostics). Human CXCL1 quantification was performed by ELISA measurements according to the manufacturer’s guidelines (DY275; R&D Systems).
Schloesser D., Lindenthal L., Sauer J., Chung K.J., Chavakis T., Griesser E., Baskaran P., Maier-Habelsberger U., Fundel-Clemens K., Schlotthauer I., Watson C.K., Swee L.K., Igney F., Park J.E., Huber-Lang M.S., Thomas M.J., El Kasmi K.C, & Murray P.J. (2022). Senescent cells suppress macrophage-mediated corpse removal via upregulation of the CD47-QPCT/L axis. The Journal of Cell Biology, 222(2), e202207097.
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5

Zymosan and Curdlan Induced Cytokine Secretion

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Cells were seeded in 12 well plates at a density of 7.5×105 (SW480) cells/well. After overnight incubation, the medium was replaced with fresh medium and Zymosan (tlrl-zyn, InvivoGen, San Diego, CA) or Curdlan (C7821 Sigma-Aldrich, St. Louis, MO). To avoid possible contaminating with endotoxin, we used endotoxin-free (<0.001 EU) Zymosan or Curdlan. Where indicated the SYK inhibitors [3-(1-methyl-1H-indol-3-yl-methylene)-2-oxo-2,3-dihydro-1H-indole-5-sulfonamide], (*574711, CAS 622387–85-3 Calbiochem, Merck-Millipore, Darmstadt, Germany) and R406 (Invivogen), were added 1 hour prior to the addition of Zymosan. All treatments were performed in duplicates/triplicate. After 20 hours, supernatants were collected, and maintained at −70ºC until analysis. IL-8, CCL2, CXCL1 and GM-CSF concentrations were determined using ELISA (DY208, DY271, DY215 and DY275 respectively from R&D systems) according to the manufacturer’s instructions.
Wang L., Aschenbrenner D., Zeng Z., Cao X., Mayr D., Mehta M., Capitani M., Warner N., Pan J., Wang L., Li Q., Zuo T., Cohen-Kedar S., Lu J., Ardy R.C., Mulder D.J., Dissanayake D., Peng K., Huang Z., Li X., Wang Y., Wang X., Li S., Bullers S., Gammage A.N., Warnatz K., Schiefer A.I., Krivan G., Goda V., Kahr W.H., Lemaire M., Lu C.Y., Siddiqui I., Surette M.G., Kotlarz D., Engelhardt K.R., Griffin H.R., Rottapel R., Decaluwe H., Laxer R.M., Proietti M., Hambleton S., Elcombe S., Guo C.H., Grimbacher B., Dotan I., Ng S.C., Freeman S.A., Snapper S.B., Klein C., Boztug K., Huang Y., Li D., Uhlig H.H, & Muise A.M. (2021). Gain-of-function variants in SYK cause immune dysregulation and systemic inflammation in humans and mice. Nature genetics, 53(4), 500-510.
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6

Zymosan and Curdlan Induced Cytokine Secretion

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Cells were seeded in 12 well plates at a density of 7.5×105 (SW480) cells/well. After overnight incubation, the medium was replaced with fresh medium and Zymosan (tlrl-zyn, InvivoGen, San Diego, CA) or Curdlan (C7821 Sigma-Aldrich, St. Louis, MO). To avoid possible contaminating with endotoxin, we used endotoxin-free (<0.001 EU) Zymosan or Curdlan. Where indicated the SYK inhibitors [3-(1-methyl-1H-indol-3-yl-methylene)-2-oxo-2,3-dihydro-1H-indole-5-sulfonamide], (*574711, CAS 622387–85-3 Calbiochem, Merck-Millipore, Darmstadt, Germany) and R406 (Invivogen), were added 1 hour prior to the addition of Zymosan. All treatments were performed in duplicates/triplicate. After 20 hours, supernatants were collected, and maintained at −70ºC until analysis. IL-8, CCL2, CXCL1 and GM-CSF concentrations were determined using ELISA (DY208, DY271, DY215 and DY275 respectively from R&D systems) according to the manufacturer’s instructions.
Wang L., Aschenbrenner D., Zeng Z., Cao X., Mayr D., Mehta M., Capitani M., Warner N., Pan J., Wang L., Li Q., Zuo T., Cohen-Kedar S., Lu J., Ardy R.C., Mulder D.J., Dissanayake D., Peng K., Huang Z., Li X., Wang Y., Wang X., Li S., Bullers S., Gammage A.N., Warnatz K., Schiefer A.I., Krivan G., Goda V., Kahr W.H., Lemaire M., Lu C.Y., Siddiqui I., Surette M.G., Kotlarz D., Engelhardt K.R., Griffin H.R., Rottapel R., Decaluwe H., Laxer R.M., Proietti M., Hambleton S., Elcombe S., Guo C.H., Grimbacher B., Dotan I., Ng S.C., Freeman S.A., Snapper S.B., Klein C., Boztug K., Huang Y., Li D., Uhlig H.H, & Muise A.M. (2021). Gain-of-function variants in SYK cause immune dysregulation and systemic inflammation in humans and mice. Nature genetics, 53(4), 500-510.
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