Opteia kit

Manufactured by BD

Sourced in United States, Germany, Italy

The OptEIA kit is a laboratory equipment product that provides a standardized platform for enzyme-linked immunosorbent assay (ELISA) analysis. The kit includes the necessary components, such as pre-coated microtiter plates, detection reagents, and buffers, to perform quantitative or qualitative measurements of target analytes in biological samples.

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BD OptEIA (350 citations) BD OptEIA ELISA kits (114 citations) BD OptEIA human ELISA kits (24 citations) BD OptEIA Human C3a ELISA Kit (7 citations) OptEIA mouse IgE ELISA kit (4 citations) IL-10 OptEIA ELISA kit (4 citations) BD OptEIA mouse ELISA kits (3 citations) OptEIA TNF-α ELISA kits (3 citations) IL-6 (BD OptEIA) ELISA kits (3 citations) Human TNF-α OptEIA ELISA kit (3 citations)

130 protocols using opteia kit

Immune Cell Cytokine and MMP3 Assay

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500 μl PB was diluted (1:1) with complete culture medium and dispensed in 24-well plates. Cells were stimulated with following ligands (in a 5% CO₂ incubator at 37 °C) for 24 h: LPS (2.5 μg/ml), PG (5 μg/ml), MRP8 (5 μg/ml), TNC (10 μg/ml). Unstimulated cells served as the baseline. In the culture supernatants, levels of TNF (BD OptEIA Kit, CA, USA), IL-6 (BD OptEIA Kit, CA, USA) and MMP3 (R&D systems, MN, USA) were measured by enzyme linked immunosorbent assay (ELISA), as per the manufacturer’s instructions. The minimum detection limit was 7.8 pg/ml for TNF, 4.7 pg/ml for IL-6 and 31.3 pg/ml for MMP3.

Bhattacharya S., Misra R, & Aggarwal A. (2020). Patients with enthesitis related arthritis show similar monocyte function pattern as seen in adult axial spondyloarthropathy. Pediatric Rheumatology Online Journal, 18, 6.

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Cytokine Response to Hemocyanin Treatments

Cited 1 time

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J774.2 cells (5 × 10⁵) were incubated with native, dissociated, and N-deglycosylated hemocyanins (500 μg/ml) in serum-free culture medium for 24 h at 37 °C as described by Jiménez et al. (30 (link)). As a positive control, cells were incubated with LPS from E. coli (Enzo Life Sciences) (100 ng/ml). As a negative control, cells were incubated with culture medium without hemocyanin. Culture supernatants were collected and quantified by ELISA using BD OptEIA^TM kits (BD Biosciences) to detect IL-6, IL-12p40, and TNFα, according to the manufacturer's instructions.

Salazar M.L., Jiménez J.M., Villar J., Rivera M., Báez M., Manubens A, & Becker M.I. (2019). N-Glycosylation of mollusk hemocyanins contributes to their structural stability and immunomodulatory properties in mammals. The Journal of Biological Chemistry, 294(51), 19546-19564.

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Quantifying Inflammatory Markers in Mice

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Insulin levels were quantified using an Ultrasensitive Mouse Insulin ELISA (Mercodia, Uppsala, Sweden). Leptin and resistin levels were measured using: Mouse/Rat Leptin and Mouse Resistin Quantikine ELISA kits (R&D Systems, Minneapolis, MN, USA). IL-1β and IL-6 dosages were carried out with ELISA Ready-SET-Go kits (eBioscience/Affymetrix, ThermoFisher Scientific). IL-10 levels were determined using BD OptEIA^TM kits (BD Biosciences, Franklin Lakes, NJ, USA).

Ayari A., Rosa-Calatrava M., Lancel S., Barthelemy J., Pizzorno A., Mayeuf-Louchart A., Baron M., Hot D., Deruyter L., Soulard D., Julien T., Faveeuw C., Molendi-Coste O., Dombrowicz D., Sedano L., Sencio V., Le Goffic R., Trottein F, & Wolowczuk I. (2020). Influenza infection rewires energy metabolism and induces browning features in adipose cells and tissues. Communications Biology, 3, 237.

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Quantifying Cytokine Profiles in Tissue

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Cytokines levels were determined from colon and/or tumor samples. Tissues were collected, weighed and immediately homogenized in PBS in the presence of protease inhibitors (Roche) using a tissue homogenizer (Polytron System PT 1200E). The material was then centrifuged at 6000 rpm for 15 min at 4 °C and the supernatant collected, aliquoted and stored at -80 °C until the time of use. The concentrations of TNF-α, IL-6, IL-12 (p70), IL-17, IFN-γ, IL-1 β, IL-10, TGF-β, IL-23 were determined by ELISA BD OptEIATM kits (BD Biosciences Pharmingen) or DuoSet (R&D Systems). The protocol was performed according to the manufacturers’ instructions. Cytokine concentrations were determined with reference to the linear regression line obtained with the serial dilution data of each recombinant mouse cytokine.

Silveira D.S., Veronez L.C., Lopes-Júnior L.C., Anatriello E., Brunaldi M.O, & Pereira-da-Silva G. (2020). Lactobacillus bulgaricus inhibits colitis-associated cancer via a negative regulation of intestinal inflammation in azoxymethane/dextran sodium sulfate model. World Journal of Gastroenterology, 26(43), 6782-6794.

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Measuring Secreted Inflammatory Factors

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CCL20 and eNAMPT were measured using Duoset kits (R&D Systems), whereas CCL5, CCL2, and CXCL8 levels were measured with OptEIA^TM kits (BD Pharmingen, Milan, Italy), in cell-free supernatants (sups) from resting or stimulated keratinocyte, fibroblast and/or endothelial cultures, according to the manufacturer’s protocols. The plates were analyzed in an ELISA reader mod.3550 UV BioRad. Results are graphed as pg or ng/10⁶ cells ± SD.

Mercurio L., Morelli M., Scarponi C., Scaglione G.L., Pallotta S., Avitabile D., Albanesi C, & Madonna S. (2021). Enhanced NAMPT-Mediated NAD Salvage Pathway Contributes to Psoriasis Pathogenesis by Amplifying Epithelial Auto-Inflammatory Circuits. International Journal of Molecular Sciences, 22(13), 6860.

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Quantifying Keratinocyte Secreted Factors

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IGFBP2, IL-1β and CCL20 were measured with Duoset kits (R&D Systems), whereas and IL-6, CXCL8 and CCL2 levels were measured with OptEIATM kits (BD Pharmingen, Milan, Italy), in cell-free supernatants (sups) from resting or stimulated keratinocyte cultures, according to the manufacturer’s protocols. The plates were analysed in an ELISA reader mod.3550 UV Bio-Rad. Results are graphed as pg or ng/10⁶ cells ± SD.

Mercurio L., Lulli D., Mascia F., Dellambra E., Scarponi C., Morelli M., Valente C., Carbone M.L., Pallotta S., Girolomoni G., Albanesi C., Pastore S, & Madonna S. (2020). Intracellular Insulin-like growth factor binding protein 2 (IGFBP2) contributes to the senescence of keratinocytes in psoriasis by stabilizing cytoplasmic p21. Aging (Albany NY), 12(8), 6823-6851.

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Evaluation of Inflammatory Response to μDH Particles

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C57BL/6 mice were randomly divided into four groups (n=3) and injected
intravenously with 50 μl of PBS solution containing different
concentrations of μDH particles (0.1, 1, and 5 mg/mL). At 24 h
post-administration, blood was collected from mice through cardiac puncture
after euthanization. A blood sample taken from the untreated mice was included
as a control. Inflammatory cytokines (IL-1β and TNF-α) in the
blood were measured using BD OptEIA kits (BD Biosciences, San Jose, CA)
following the manufacturer’s protocol.

Wang J., Cooper R.C., He H., Li B, & Yang H. (2018). Polyamidoamine Dendrimer Microgels: Hierarchical Arrangement of Dendrimers into Micrometer Domains with Expanded Structural Features for Programmable Drug Delivery and Release. Macromolecules, 51(15), 6111-6118.

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Quantifying Cytokine Levels by ELISA

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For the quantification of cytokines by ELISA (Enzyme-Linked ImmunoSorbent Assay), BMDMs were plated in 24-well plates (5 × 105 cells/well) and pretreated with 100 ng/ml LPS (Escherichia coli 055: B5 LPS, Sigma) or Pam3Cys for 4 h and then infected with L. amazonensis (MOI 10). The cytokine IL-1β present in the supernatant was measured by ELISA using the BD OptEIA kits (BD Biosciences), according to instructions provided by the manufacturer. The OD values were read on the SpectraMax i3 system (Molecular Devices).

de Sá K.S., Amaral L.A., Rodrigues T.S., Ishimoto A.Y., de Andrade W.A., de Almeida L., Freitas-Castro F., Batah S.S., Oliveira S.C., Pastorello M.T., Fabro A.T, & Zamboni D.S. (2023). Gasdermin-D activation promotes NLRP3 activation and host resistance to Leishmania infection. Nature Communications, 14, 1049.

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Cytokine Expression in Immunized Mice

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For estimation of cytokines, three mice from immunized group were randomly selected and sacrificed and their splenocytes were removed aseptically and resuspended in RPMI-1640 media with 10% FBS. The splenocytes from un-immunized mice group were used as negative controls. The spleen cells were seeded in a 24-well plate at a density of 1 × 10⁶ cells/well. The isolated splenocytes cultures were stimulated with 5 μg/mL of rBoNT/E-LC. These cells were incubated at 37 °C in CO₂ incubator (5% CO₂) for 48 h. Supernatants were collected after centrifugation (1,500 × g) and stored at – 80 °C until analyzed for cytokines expression. The expression levels of different cytokines i.e. IFN-γ, TNF-α, IL-2, IL-4 and IL-10 were determined by ELISA using BD OptEIA Kits, according to the manufacturer's protocol. The expression of cytokines was calculated using standard curves generated with known concentrations of recombinant cytokines and expressed in picograms per milliliter (pg/mL).

Sonkar P., Chauhan V., Chauhan R., Saxena N, & Dhaked R.K. (2020). Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E. Scientific Reports, 10, 13932.

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ELISA Analysis of Cell Supernatants

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After cultivation, cell-free supernatants were analyzed by Enzyme-linked Immunosorbent Assay (ELISA) using BD OptEIA™ kits (BD Biosciences) according to the manufacturer’s protocols.

Meyer J.E., Loff S., Dietrich J., Spehr J., Jurado Jiménez G., von Bonin M., Ehninger G., Cartellieri M, & Ehninger A. (2021). Evaluation of switch-mediated costimulation in trans on universal CAR-T cells (UniCAR) targeting CD123-positive AML. Oncoimmunology, 10(1), 1945804.

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