El800 elisa reader

Manufactured by Agilent Technologies

Sourced in United States

The EL800 ELISA reader is a compact and easy-to-use microplate reader designed for absorbance-based ELISA assays. It features a high-quality optical system and a user-friendly interface, providing reliable results for a variety of applications.

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Lab products found in correlation

Anti bovine igg, by Merck Group (1 mentions) T0440, by Merck Group (1 mentions) Instantone elisa, by Thermo Fisher Scientific (1 mentions) Ripa lysis buffer kit, by Santa Cruz Biotechnology (1 mentions) 96 well immunoplate, by Thermo Fisher Scientific (1 mentions) Spss version 12, by IBM (1 mentions) Il 1β, by Cell Signaling Technology (1 mentions) Interleukin 6 (il 6), by Abcam (1 mentions)

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ELISA reader (447 citations) EL800 (239 citations) EL800 reader (5 citations)

10 protocols using el800 elisa reader

ESAT-6/MPB70/MPB83 Chimera ELISA

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The wells of Costar 3590 polystyrene 96-well plates (Corning, Corning, NY, U.S.A.) were adsorbed with the recombinant chimera of ESAT-6/MPB70/MPB83 [22 (link)] in carbonate-bicarbonate buffer, pH 9.6, for 60 min at 37°C. The plates were then blocked with 100 µl/well of phosphate buffer saline with 0.1% Tween 20 (PBST)
with 5% skim milk for 60 min at 37°C. After five washes with PBST, 100 µl/well of the control and test sera diluted to 1:600 in PBST with 2% skim milk were incubated for 60
min at 37°C. The plates were washed five times with PBST. Next, 100 µl of anti-bovine IgG (whole molecule) horseradish peroxidase conjugate (A8917, Sigma, St. Louis, MO,
U.S.A.) (dilution: 1:80,000 in PBST) were added to each well. The plates were incubated for 30 min at 37°C, washed five times, and 50 µl/well of chromogen/substrate
tetramethylbenzidine (T0440, Sigma) was added. The reactions were stopped with 2.5 N of H₂SO₄, and the results were read in an EL-800 ELISA reader (BioTek Instruments,
Winooski, VT, U.S.A.) with a 450 nm filter.

de SOUZA I.I., RODRIGUES R.D., GONÇALVES JORGE K.S., SILVA M.R., LILENBAUM W., VIDAL C.E., ETGES R.N., KOSTOVIC M, & ARAÚJO F.R. (2018). ELISA using a recombinant chimera of ESAT-6/MPB70/MPB83 for Mycobacterium bovis diagnosis in naturally infected cattle. The Journal of Veterinary Medical Science, 81(1), 9-14.

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NF-κB Pathway Activation in HCMV-Infected Macrophages

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Whole cell extracts were prepared from 10×10⁶ mock-infected or HCMV-infected macrophages cultured in the presence or absence of LPS (1 µg/mL) for 15 min at 37°C using the RIPA Lysis Buffer Kit (Santa Cruz Biotechnology, Santa Cruz, CA). Phosphorylated NF-κB p65 and IκBα were analyzed using the InstantOne ELISA (eBioscience), which detects total and phosphorylated NF-κB p65 and IκBα attached to consensus binding sites in a 96-well plate using the TMB colorimetric substrate and optical density at 450 nm (EL 800 ELISA reader, Biotek Instruments, Inc., Winooski, VT). Data is presented as phosphorylated NF-κB p65 and IκBα at OD 450 nm/10 µg protein.

Smith P.D., Shimamura M., Musgrove L., Dennis E.A., Bimczok D., Novak L., Ballestas M., Fenton A., Dandekar S., Britt W.J, & Smythies L.E. (2014). Cytomegalovirus Enhances Macrophage TLR Expression and MyD88-mediated Signal Transduction to Potentiate Inducible Inflammatory Responses. Journal of immunology (Baltimore, Md. : 1950), 193(11), 5604-5612.

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Quantifying MUC5B and MUC8 Protein Levels

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Protein levels for MUC5B and MUC8 were determined by enzyme-linked immunosorbent assay (ELISA). Sample of cell lysates from NCI-H292 cells was incubated at 40℃ in a 96-well plate until dry. Plates were then washed three times with PBS, blocked with 2% bovine serum albumin for 1 hour at room temperature, washed again three times with PBS, and incubated with primary antibody for MUC8 or MUC5B diluted at 1:200 with PBS containing 0.05% tween 20 for 1 hour. Wells were treated by dispensation of HRP-conjugated secondary antibody into each well. After 4 hours, color was developed using 3,3', 5,5'-tetramethylbenzidine peroxidase solution and stopped with 2N-H₂SO₄. Optical density measurements were performed using an EL800 ELISA reader (BIO-TEK Instruments, Winooski, VT, USA) at 450 nm.

Bae C.H., Jeon B.S., Choi Y.S., Song S.Y, & Kim Y.D. (2014). Delphinidin Inhibits LPS-Induced MUC8 and MUC5B Expression Through Toll-like Receptor 4-Mediated ERK1/2 and p38 MAPK in Human Airway Epithelial Cells. Clinical and Experimental Otorhinolaryngology, 7(3), 198-204.

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Quantification of MUC5AC Protein

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NCI-H292 cells and human nasal epithelial cells were prepared and treated with recombinant nCLU as described above. Following recombinant nCLU incubation for 24 hours, MUC5AC protein level was determined using an enzyme-linked immunosorbent assay (ELISA). The color was developed using 3,3´, 5,5´-tetramethylbenzidine peroxidase solution and stopped with 2N-H₂SO₄. Optical density measurements were performed using an EL800 ELISA reader (BioTek Instruments, Winooski, VT, USA) at 450 nm. The results were expressed as percent of baseline control.

Bae C.H., Na H.G., Choi Y.S., Song S.Y, & Kim Y.D. (2018). Clusterin Induces MUC5AC Expression via Activation of NF-κB in Human Airway Epithelial Cells. Clinical and Experimental Otorhinolaryngology, 11(2), 124-132.

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MUC5AC Protein Quantification in Epithelial Cells

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NCI-H292 cells and human nasal epithelial cells were treated with e-cigarette vapor with or without nicotine, as described above. After further incubation for 24 hours, MUC5AC protein levels were determined using enzyme-linked immunosorbent assay (ELISA). Lysates sample from epithelial cells were incubated overnight at 4°C, then plates were blocked with 2% bovine serum albumin (BSA) and incubated with the primary antibody in PBS containing 0.05% Tween 20 for 1 hour. Wells were washed and treated with the secondary antibody for 1 hour. Color was developed using 3,3ʹ,5,5ʹ-tetramethylbenzidine peroxidase solution, and the reaction was terminated with 2N H₂SO₄. Optical density measurements were obtained using an EL800 ELISA reader (BioTek Instruments, Winooski, VT, USA) at 450 nm. Results are expressed as fold increase above baseline controls.

Song S.Y., Na H.G., Kwak S.Y., Choi Y.S., Bae C.H, & Kim Y.D. (2020). Changes in Mucin Production in Human Airway Epithelial Cells After Exposure to Electronic Cigarette Vapor With or Without Nicotine. Clinical and Experimental Otorhinolaryngology, 14(3), 303-311.

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Indirect ELISA for TcCruzipain Detection

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For indirect ELISA, recombinant TcCruzipain (0.15 μg/well) was adsorbed onto 96-well immunoplates (Nunc, Roskilde, Denmark) by incubation overnight at 4°C with sensitizing buffer (0.05 M sodium carbonate and sodium bicarbonate, pH 9.6). The plates were then blocked by incubation for 1 h with 5% nonfat milk powder in PBS supplemented with 0.01% Tween 20 (PBS-T). The hybridoma supernatants were added to the immunoplates and incubated for 1 h at 37°C. The plates were washed five times with PBS-T and incubated for 1 h at 37°C with HRP-conjugated goat anti-mouse IgG (1 : 4,000). The plates were then washed five times with PBS-T and immunoreactivity was visualized with the SureBlue TMB Substrate, with optical density (OD) being read at 450 nm in an EL800 ELISA reader (BioTek, Winooski, VT, USA). Only OD values higher than 0.300 were considered positive.

Batista C.M., Medeiros L.C., Eger I, & Soares M.J. (2014). mAb CZP-315.D9: An Antirecombinant Cruzipain Monoclonal Antibody That Specifically Labels the Reservosomes of Trypanosoma cruzi Epimastigotes. BioMed Research International, 2014, 714749.

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Quantification of Inflammatory Cytokines

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Statistical analyses were performed using SPSS, version 12.0 software (SPSS, Chicago, IL, USA). Data were expressed as the mean ± standard deviation (SD). Comparisons were performed using the unpaired t-test or Kruskal-Wallis test followed by a Mann-Whitney test. For all tests, a p value < 0.05 was considered statistically significant.
were blocked with 2 % bovine serum albumin (BSA) for 1 h and incubated with the following primary antibodies: rabbit anti-MUC5AC (H-160) (sc-20118; Santa Cruz Biotechnology, USA; 1:200 dilution), IL-1β (#12242; Cell Signaling Technology, USA; 1:1000 dilution), IL-6 (ab6672; Abcam, Cambridge, UK; 1:1000 dilution), and IL-8 (M801; Thermo Scientific, IL, USA; 1:1000 dilution) in PBS containing 0.05 % Tween 20 for 1 h. Then, they were incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody. After 1 h, colour was developed using 3,3' ,5,5'-tetramethylbenzidine peroxidase solution, and the reaction was stopped using 2 N H 2 SO 4 . Optical density was measured using an EL800 ELISA reader at 450 nm (BIO-TEK Instruments, Winooski, VT, USA). Results were expressed as fold increase from the baseline control.

Kwak S., Choi Y.S., Na H.G., Bae C.H., Song S.Y, & Kim Y.D. (2020). Fipronil upregulates inflammatory cytokines and MUC5AC expression in human nasal epithelial cells. Rhinology, 58(1).

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Antimicrobial Evaluation of Soursop Kombucha

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The antimicrobial activity of soursop kombucha samples was evaluated against pathogenic bacteria including Escherichia coli 0157:H7 and Staphylococcus aureus ATCC6538 using the microtiter plate assay (Muhialdin et al., 2020b (link)). The sample (100 μl) was pipetted into the wells of microtiter plates and mixed with 100 μl of nutrient broth containing 10⁶ CFU/ml. The nutrient broth (200 μl) containing the tested bacteria was pipetted into the wells of microtiter plates as control. The plates were incubated at 37°C for 24 h. The growth of inhibition of targeted bacteria was measured at OD₆₀₀ using the BioTek EL × 800 ELISA reader. The percentage growth of E. coli and S. aureus was calculated according to the following formula:

Tan W.C., Muhialdin B.J, & Meor Hussin A.S. (2020). Influence of Storage Conditions on the Quality, Metabolites, and Biological Activity of Soursop (Annona muricata. L.) Kombucha. Frontiers in Microbiology, 11, 603481.

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MUC5AC Quantification in NCI-H292 Cells

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After incubation for 24 h, DEP was treated to the NCI–H292 cells as previously described. The protein levels of MUC5AC were evaluated by ELISA. Supernatants of NCI–H292 cells were collected. The samples diluted with PBS were transferred to an F96 certified MaxiSorp Nunc-Immuno Plate (Fisher Scientific, Lenexa, KS, USA) and incubated overnight at 4 °C. Subsequently blocked with 2% bovine serum albumin (BSA) for 1 h and incubated with 1:200 dilution of rabbit anti-MUC5AC antibody (#sc-20118; Santa Cruz Biotechnology, USA), the samples were incubated with an HRP-conjugated secondary antibody. After 1 h, we incubated each well with 3,3′,5,5′-tetramethylbenzidine (TMB) peroxidase substrate solution, terminating the reaction with 2 N sulfuric acid (H₂SO₄). Using EL800 ELISA reader by BIO-TEK Instruments (Winooski, VT, USA), we measured optical density at 450 nm and indicated the data as a fold increase to the control.

Jo S., Na H.G., Choi Y.S., Bae C.H., Song S.Y, & Kim Y.D. (2022). Saponin attenuates diesel exhaust particle (DEP)-induced MUC5AC expression and pro-inflammatory cytokine upregulation via TLR4/TRIF/NF-κB signaling pathway in airway epithelium and ovalbumin (OVA)-sensitized mice. Journal of Ginseng Research, 46(6), 801-808.

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Measuring Mucin Secretion in Airway Epithelial Cells

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We treated the NCI-H292 and HNEpC lines with eotaxin-2/3, as previously described. MUC5AC protein levels were determined using ELISA. Cell lysates were scraped off the plate with the radioimmunoprecipitation assay (RIPA) buffer (200 μL) and diluted with PBS solution. They were then transferred to the F96 Cert. Maxisorp Nunc-Immuno Plate (Thermo Fisher Scientific). After overnight incubation at 4°C, samples were blocked with 2% bovine serum albumin (BSA) and incubated with the primary antibody in PBS containing 0.05% Tween 20 for 1 h. HRPconjugated secondary antibodies were applied to each well. After 4 h, they were stained with the 3, 3′, 5, 5′tetramethylbenzidine peroxidase solution that was stopped with 2 N H 2 SO 4 . Using an EL800 ELISA reader (BioTek Instruments, Winooski, VT, USA), we acquired the data via optical density measurement at 450 nm, indicated as percentages of the baseline control.

Jo S., Na H.G., Choi Y.S., Bae C.H., Song S.Y, & Kim Y.D. (2023). C-C Motif Chemokine Receptor 3-Mediated Extracellular Signal-Regulated Kinase 1/2 and p38 Mitogen-Activated Protein Kinase Signaling: Promising Targets for Human Airway Epithelial Mucin 5AC Induction by Eotaxin-2 and Eotaxin-3. International archives of allergy and immunology, 184(9).

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