Dla50

Manufactured by R&D Systems

Sourced in United Kingdom

The DLA50 is a laboratory equipment product designed for performing cell culture assays. It provides precise control over environmental conditions, such as temperature and humidity, to ensure optimal cell growth and experimental consistency.

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Lab products found in correlation

7 protocols using dla50

Biomarker Quantification in Plasma

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Commercially available ELISA kits were used to measure the concentrations or activity of IFNβ (DIFNB0 or 42400–1, R&D Systems), F3 (DCF300, R&D Systems; ab214091, Abcam), fibrin (MBS265263 or MBS706338, MyBioSource), D-dimer (ab196269, Abcam; MBS723281, MyBioSource), TNF (DTA00D, R&D Systems), IL1A (DLA50, R&D Systems), IL1B (DLB50, R&D Systems), IL6 (D6050, R&D Systems), and HMGB1 (ST51011, IBL International) in the indicated samples. Measurement of GPT/ALT and BUN in the plasma was performed using an IDEXX Catalyst Dx Chemistry Analyzer. PT, APTT, and fibrinogen were measured in an automated coagulometer (Sysmex CA-7000). Platelet count was measured using an IDEXX ProCyte Dx Hematology Analyzer.

Zhang H., Zeng L., Xie M., Liu J., Zhou B., Wu R., Cao L., Kroemer G., Wang H., Billiar T.R., Zeh H.J., Kang R., Jiang J., Yu Y, & Tang D. (2020). TMEM173 Drives Lethal Coagulation in Sepsis. Cell host & microbe, 27(4), 556-570.e6.

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Rhodosorus marinus Extract Effects on NGF and IL-1a

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Cytotoxicity tests were performed to confirm that the Rhodosorus marinus extract was not toxic on NHEK and NHA (data not shown). Pooled normal human epidermal keratinocytes (adult, from Lonza) were seeded in 12-well plates at 100,000 cells per well. After 48 h, cells were washed in PBS (Gibco^TM, Thermo Fischer Scientific, Illkirch, France) and the culture medium was replaced by KGM (Keratinocytes Growth Medium, Lonza, Basel, Switzerland) without hydrocortisone, an anti-inflammatory molecule that can influence the PMA effect and consequently the secretion of NGF and IL-1a. After 24 h, cells were prestimulated with dexamethasone (10 µM) or Rhodosorus marinus extract at 1% and 3% for 2 h at 37 °C. Cells were then stimulated with PMA (phorbol myristate acetate, Sigma) at 10 ng/mL. After 48 h of stimulation at 37 °C, supernatant was collected, the released NGF was quantified by ELISA (Abcam, Cambridge, UK), and the secretion of IL-1a was quantified by ELISA from R&D system (DLA50). For each ELISA, the experimental procedure recommended by suppliers was used and the measurements were performed using a TECAN microplate reader.

Scandolera A., Hubert J., Humeau A., Lambert C., De Bizemont A., Winkel C., Kaouas A., Renault J.H., Nuzillard J.M, & Reynaud R. (2018). GABA and GABA-Alanine from the Red Microalgae Rhodosorus marinus Exhibit a Significant Neuro-Soothing Activity through Inhibition of Neuro-Inflammation Mediators and Positive Regulation of TRPV1-Related Skin Sensitization. Marine Drugs, 16(3), 96.

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Quantification of Protein Biomarkers

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Enzyme‐linked immunosorbent assay (ELISA) kits were used to detect the concentrations of human FGF19 (DF1900, R&D Systems), IL‐1α (DLA50, R&D Systems), IL‐1β (DLB50, R&D Systems) and complement C5a (ab193695, Abcam) in culture supernatants or serum as described.^[³⁶^] The absorbance values could be read on a microplate reader (Molecular Devices) at a wavelength of 450 nm, within 30 min. The standard curve was used to convert absorbance values to protein concentrations.

Li C., Chen T., Liu J., Wang Y., Zhang C., Guo L., Shi D., Zhang T., Wang X, & Li J. (2023). FGF19‐Induced Inflammatory CAF Promoted Neutrophil Extracellular Trap Formation in the Liver Metastasis of Colorectal Cancer. Advanced Science, 10(24), 2302613.

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Quantification of Cytokines and HSPs

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Enzyme-linked immunosorbent assay (ELISA) was used for quantitative determination of IL-1α (#DLA50, R&D Systems, Bio-Techne), HMGB1 (#NBP2-62766, Novus Biologicals, supplied by Bio-Techne) and HSP90α (#NBP2-76448, Novus Biologicals) in ACS conditioned medium. Protocols for all ELISAs adhered to the manufacturer’s instructions and data was obtained using a FLUOstar OPTIMA plate reader by spectrophotometric detection at 450 nm as previously (Dunnill et al., 2020 (link)).

Peake M., Dunnill C., Ibraheem K., Smith A., Clarke D.J, & Georgopoulos N.T. (2024). A novel method for the establishment of autologous skin cell suspensions: characterisation of cellular sub-populations, epidermal stem cell content and wound response-enhancing biological properties. Frontiers in Bioengineering and Biotechnology, 12, 1386896.

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Quantitation of Biomarkers in Clinical Samples

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IL1-α, IL-8, and VEGF-A in cell supernatant were analyzed using Quantikine ELISA kits (R&D Systems; DLA50, D8000C, DVE00). Human GRO-beta and GRO-gamma ELISA Construction kits (Antigenix America Inc.; RHF810CK, RHF820CKP) were used to measure the supernatant and plasma levels of CXCL2 and CXCL3, respectively. IL-1α, IL-8, and VEGF-A in human plasma were analyzed using custom V-PLEX Assays (Meso Scale Discovery, Maryland, USA; K151RBD-2, K151RAG-1, K151RHD-1). Patient blood samples were obtained after multicenter ethics approval (09/H1102/107 and 14/YH/0090) as described previously [16 (link), 22 (link)]. Blood was centrifuged within 1 h of collection at 2500 rpm in a lithium-heparin containing tube; the plasma was then removed, aliquoted, and stored in a − 70 °C freezer prior to thawing for ELISA analysis. ELISAs were carried out according to the manufacturer’s instructions.

Phillips M.M., Pavlyk I., Allen M., Ghazaly E., Cutts R., Carpentier J., Berry J.S., Nattress C., Feng S., Hallden G., Chelala C., Bomalaski J., Steele J., Sheaff M., Balkwill F, & Szlosarek P.W. (2023). A role for macrophages under cytokine control in mediating resistance to ADI-PEG20 (pegargiminase) in ASS1-deficient mesothelioma. Pharmacological Reports, 75(3), 570-584.

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Cytokine ELISA Quantification in Plasma and Media

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For ELISA of plasma, blood was harvested via cardiac puncture from anesthetized mice at end point and plasma separated by centrifugation. For ELISA of media, conditioned media was harvested as described above. ELISA assays used were IL-1α (DLA50; R and D Systems), IL-1β (DLB50; R and D Systems) and IL-6 (M6000B; R and D Systems).

Somerville T.D., Biffi G., Daßler-Plenker J., Hur S.K., He X.Y., Vance K.E., Miyabayashi K., Xu Y., Maia-Silva D., Klingbeil O., Demerdash O.E., Preall J.B., Hollingsworth M.A., Egeblad M., Tuveson D.A, & Vakoc C.R. (2020). Squamous trans-differentiation of pancreatic cancer cells promotes stromal inflammation. eLife, 9, e53381.

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Quantification of Cytokines in HCLE Cell Conditioned Media

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To quantify cytokines in CM obtained from HCLE cells, two different approaches were taken. First, we performed ELISA assays for TGFβ1, IL-1α, and IL-1RA on CM obtained from three replicate sets of control and CM from MMC treated cells grown in Low GF HCLE media. Kits were obtained from R&D Systems (human TGFβ1: DB100C; human IL-1α/IL-1F1: DLA50, human IL1RA/IL-1F3: DRA008) and assays performed as per the manufacter’s instructions. For the ELISA assay of TGFβ1, activation was performed by treating each replicate CM using R&D Systems Sample Activation kit (DY010). ELISA samples were read using a Tecan proM200 plate reader equipped with Magellen software.
We also performed Luminex assays on HCLE-CMC and CMM and IMDM HCLE-CMC and CMM. A 96 well Invitrogen/ProcartaPlex multiplex immunoassay plate was custom ordered from ThermoFisher to allow us to simultaneously detect the following human cytokines: IL-6, IL-1α, IL-1β, TNFα, IL-17F, IL-12p70, IL-1RA, and IL-23. Cytokine concentrations in CM samples were read using a Luminex Instrument (xPONENT 3.1, Luminex 100 IS Version 2.3, Austin, TX) equipped with Millipex analyst software version 5.1 (Millipore, Burlington, MA).

Pal-Ghosh S., Karpinski B.A., Majumdar H.D., Ghosh T., Thomasian J., Brooks S.R., Sawaya A.P., Morasso M.I., Scholand K.K., de Paiva C.S., Galletti J.G, & Stepp M.A. (2022). Molecular mechanisms regulating wound repair: evidence for paracrine signaling from corneal epithelial cells to fibroblasts and immune cells following transient epithelial cell treatment with Mitomycin C. Experimental eye research, 227, 109353.

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