IgG Aspergillus antibodies were determined in serum samples from every patient using Aspergillus fumigatus IgG ELISA (IBL-America) and the readings were made in ELISA Reader ELx808, Bio-Tek. As stated by the manufacturer, definitions for negative, equivocal and positive results were applied according to the titer levels (negative: < 8 U/mL; equivocal: 8–12 U/mL; positive: > 12 U/mL).
Elx808 elisa reader
Manufactured by Agilent Technologies
Sourced in United States
The ELx808 ELISA reader is a microplate reader designed for enzyme-linked immunosorbent assay (ELISA) applications. It provides accurate absorbance measurements across a range of wavelengths to support various ELISA protocols.
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Lab products found in correlation
Elisa plate, by Thermo Fisher Scientific (2 mentions)
Ip 10, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Scd163, by R&D Systems (1 mentions)
Scd14, by R&D Systems (1 mentions)
Maxisorb, by Thermo Fisher Scientific (1 mentions)
Quanti blue alkaline phosphatase detection medium, by InvivoGen (1 mentions)
Sba clonotyping system, by Southern Biotech (1 mentions)
Alkaline phosphatase substrate, by Bio-Rad (1 mentions)
Cellular dna fragmentation elisa kit, by Roche (1 mentions)
Wst 1, by Merck Group (1 mentions)
Wst 1, by Roche (1 mentions)
Cell counting kit, by Dojindo Laboratories (1 mentions)
18 protocols using elx808 elisa reader
1
Aspergillus IgG Antibody Detection
2
Serum Biomarkers for Bone Turnover
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Bone formation and resorption were monitored using the serum levels of P1NP and CTX, respectively. Serum samples were measured before the OVX/Sham operations (day 0) and 2 and 8 weeks after the surgery. The CTX and P1NP were determined using RATLAPS EIA and mouse P1NP EIA kits, respectively, both purchased from IDS (Tyne & Wear, UK). The results were recorded in an Elisa Reader ELx808 (Bio-Tek Instruments Inc., Winooski, VT, USA) according to the manufacturer’s instructions.
3
Quantification of Striatal Dopamine and Neuroinflammation
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Striatal DA was estimated using the Rat Dopamine ELISA kit (USCN Life Science Inc., Wuhan, China). Similarly, TNF-α was assayed using a commercial ELISA assay kit (USCN Life Science Inc., Wuhan, China). A rat IL-1βsolid-phase Sandwich ELISA (Nawah, Egypt) was used for the analysis of IL-1β. All procedures were carried out according to the manufacturers' instructions. In brief, samples (diluted 1 : 2) were added in triplicate to the microtiter plate wells coated with the antibody specific to DA or TNF-α, and the plates were incubated for 2 h at 37°C. After removing the unbound substances from each well, a biotin-conjugated antibody was added to the wells, and the plates were incubated for 1 h at 37°C. After washing, wells were incubated with an avidin-conjugated HRP for 30 min at 37°C. Finally, a stop solution was added to stop the reaction, and the color changes were quantified at 450 nm using the BioTek ELisa Reader ELx808. The results were expressed as pg/mg protein.
4
Plasma Biomarker Quantification Protocol
5
Quantifying Apoptosis with ELISA
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A Cellular DNA Fragmentation ELISA Kit (11585045001; Roche Applied Science, CH) was used to measure cytoplasmic histone-associated DNA fragments, which reflect the percentage of apoptotic cells.26 (link) This assay is based on the quantitative detection of BrdU-labelled DNA fragments. Using camptothecin (50 μM, 24 h) as a positive control, the optimal apoptotic response was detected after administration. These methods were then applied to evaluate the proportion of apoptotic cells after treatment with APS in the presence or absence of adriamycin (0.1 µg/mL) for 24 and 48 h. DNA fragmentation was determined according to the manufacturer’s instructions. Absorbance values at 450 nm were measured with an ELx808 ELISA reader (BioTek).
6
Quantifying NF-κB Activation in THP1 Cells
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THP1-Blue NF-κB cells were infected as described above and cultured for 24 h at 37 °C. Then 20 µl of culture supernatant from each well were mixed with 200 µl of QUANTI-Blue alkaline phosphatase detection medium (InvivoGen), following the manufacturer’s instructions, in a 96-well flat bottomed Maxi-Sorb (Nunc) ELISA plate and incubated for 4 h at 37 °C. Absorbance at 630 nm was measured on an ELx808 ELISA reader (BioTek).
7
Multiplex Cytokine Quantification in T Cell Cytotoxicity
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Cytokine production was measured in supernatants from the T cell–mediated cytotoxicity assays. A U-PLEX Proinflam Combo 1 (hu) multiplex assay (K15049K; MeSo Scale Discovery), custom-made combined U multiplex (MSD, K15067M-2) and R-plex assay (MIG [MSD, F210I-3] and GZMB [MSD, F213X-3]) and a Luminex assay (Milliplex MAP – human cytokine/TH17 panel [SPR1459; Millipore Sigma]) were used essentially according to manufacturer’s instructions. Perforin and GZMB release were analyzed by ELISA using the human Perforin ELISA development kit (3465-1H-6; MabTech) and the human GZMB DuoSet ELISA kit (DY2906-5; R&D Systems), according to manufacturer’s instructions. Absorbance was measured at 450 nm with an ELISA plate reader (ELx808 ELISA Reader; Biotek Instruments).
8
Serum Ig Levels in Unimmunized Mice
9
Quantifying Cellular DNA Fragmentation
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A Cellular DNA Fragmentation ELISA kit (Roche Applied Science; cat. no. 11585045001) was used according to the manufacturer's instructions. This assay is based on the quantitative detection of 5-bromo-2′-deoxyuridine (BrdU)-labeled DNA fragments. Cells were seeded at a density of 1×105 cells/well in 96-well plates. After 24 h of growth, cells were exposed to different treatments. Subsequently, 10 µM BrdU was added to each well, and the cells were cultured with BrdU for 24 h. The complexes were centrifuged at ~250 × g for 10 min at 4°C, the supernatant carefully removed. After DNA labelling, the cells were lysed in 200 µl incubation buffer and soluble DNA fragments were quantified using the Cellular DNA Fragmentation ELISA kit, according to the manufacturer's instructions. Absorbance values were detected spectrophotometrically at 450 nm using an ELx808 ELISA reader (BioTek Instruments). All experiments were performed in triplicate.
10
Antiproliferative Effects of TP and Andrographolide
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The cells were plated in a 96-well plate at a density of 1×105 cells/well for 24 h. Different concentrations of TP and andrographolide were applied either separately or in combination. Co-treatment was performed by adding the two compounds simultaneously on cells for 24 h. Pretreatment was performed by exposing the cells to andrographolide for 24 h followed by TP treatment for another 24 h. RPMI alone was added to the control cells. WST-1 (Roche) and XTT (Sigma Aldrich) reagents were used, according to the manufacturer’s guide (10 µL of WST-1 reagent was added to each well while 40 µL of XTT was added to each well), to detect cell proliferation. Cell proliferation was assessed by recording the absorbance at a wavelength of 450 nm (using a Biotek ELx808 ELISA reader), which reflects the amount of formazan dye produced by metabolically active cells.
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