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Instant elisa kit

Manufactured by Thermo Fisher Scientific
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The Instant ELISA kit is a laboratory tool used for the rapid detection and quantification of specific analytes in a sample. It is a pre-coated, ready-to-use enzyme-linked immunosorbent assay (ELISA) kit that enables efficient and accurate analysis without the need for extensive sample preparation or lengthy incubation steps.

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Lab products found in correlation

High sensitivity elisa kit, by Thermo Fisher Scientific (2 mentions) Rabbit anti caspase 3, by Cell Signaling Technology (2 mentions) Accuri c6, by BD (2 mentions) Anti cd3 anti cd28 dynabeads, by Thermo Fisher Scientific (2 mentions) Microplate reader, by Tecan (2 mentions) Synergy ht microplate reader, by Agilent Technologies (2 mentions) Penicillin streptomycin mixture, by Lonza (2 mentions) Rpmi 1640 medium, by Lonza (2 mentions) Xn 1000, by Sysmex (1 mentions) Mrx 2, by Dynex (1 mentions) Milliplex map human high sensitivity t cell panel, by Merck Group (1 mentions) Duoset elisa kit, by R&D Systems (1 mentions) Magpix system, by Merck Group (1 mentions) Erythrosin b, by Merck Group (1 mentions) 3 iodo 7 azaindole, by Merck Group (1 mentions) Lipopolysaccharide lps escherichia coli 0111 b4, by Merck Group (1 mentions) Cell proliferation reagent wst 1, by Roche (1 mentions) Prednisone, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Phorbol myristate acetate (pma), by Merck Group (1 mentions)

Close spelling variants of this product

Human Instant Enzyme-Linked Immunosorbent Assay (ELISA) kit Human MPO Instant ELISA Kit Human sCD27 Instant ELISA Kit TWEAK Human Instant ELISA Kit Mouse TNFα Instant ELISA kit Human Instant ELISA kit Human G-CSF Instant ELISA Kit ELISA Instant One kit Human IL-6 Instant ELISA Kit CRP Human Instant ELISA kit

15 protocols using instant elisa kit

1

Soluble CD27 ELISA Measurement

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Concentration of soluble CD27 in experimental supernatants was measured using an Instant ELISA kit from eBioscience (# BMS286INST). For each sample, 50μl were done in duplicate and incubated 3 hours at room temperature on rotary shaker. After washing the micro wells, 100μl TMB Substrate was added to each well and color allowed to develop for 10 minutes. Reaction was stopped by adding 100μl of Stop Solution and plate was read immediately on a SpectraMax 190 plate reader at 450nm using SoftMax Pro software program.
Yang Z.Z., Grote D.M., Xiu B., Ziesmer S.C., Price-Troska T.L., Hodge L.S., Yates D.M., Novak A.J, & Ansell S.M. (2014). TGF-β upregulates CD70 expression and induces exhaustion of effector memory T cells in B-cell non-Hodgkin’s lymphoma. Leukemia, 28(9), 1872-1884.
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2

Plasma Biomarkers in Clinical Cohort

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Plasma IL-6 concentrations were analyzed using a commercially available high sensitivity ELISA kit (eBioscience), according to the manufacturer's instructions. The limit of detection was 0.007 pg/ml. The intra- and inter-assay coefficients of variability for plasma IL-6 measurements were 10 and 12%, respectively. Plasma hsCRP concentrations were analyzed using a commercially available Instant ELISA kit (eBioscience), according to the manufacturer's instructions. The limit of detection was 3 pg/ml. The intra- and inter-assay coefficients of variability for plasma hsCRP measurements were 6 and 8%, respectively. WBC counts were obtained from a standard clinical complete blood count panel using a Sysmex XN 1000 (Sysmex).
de Punder K., Entringer S., Heim C., Deuter C.E., Otte C., Wingenfeld K, & Kuehl L.K. (2018). Inflammatory Measures in Depressed Patients With and Without a History of Adverse Childhood Experiences. Frontiers in Psychiatry, 9, 610.
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3

Apoptosis Induction by miR-214-3p and Cisplatin

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After transfection with pre-miR-214-3p (12 nM) for 48 hours, TE7 cells were treated with Cisplatin (15 μM) (clinical grade) for 24 hours and stained with fluorescein-labeled Annexin-V (FITC) and Propidium Iodide (PI) (TACS Annexin V FITC Apoptosis Detection kit, Trevigen Inc, Gaithersburg, MD). Briefly, cells were washed twice with cold PBS and spun at 300g for 5 min at RT. 106 cells were resuspended in 100μl Annexin V incubation reagent (10 μl 10× binding buffer, 2 μl PI, 1 μl Annexin V FITC and 87 μl deionized water per sample) and incubated in the dark for 15 minutes at RT. At the end of incubation, 400 μl of 1× binding buffer was added (per 100 μl reaction). Samples were then analyzed for FITC Annexin-V positive, PI negative or FITC Annexin-V positive, PI positive by flow cytometry (BD Accuri C6). For caspase activation assays, cells were exposed to Cisplatin following transfection with pre-miR-214-3p for 24 hours and cleaved caspase was measured by Western blot using rabbit anti-caspase-3 (Cell Signaling, Danvers, MA). Caspase enzyme activity was measured in triplicate for each experiment, using instant ELISA kit (eBioscience Inc., San Diego, CA), following the manufacturer's instructions.
Phatak P., Byrnes K.A., Mansour D., Liu L., Cao S., Li R., Rao J.N., Turner D.J., Wang J.Y, & Donahue J.M. (2015). Overexpression of miR-214-3p in Esophageal Squamous Cancer Cells Enhances Sensitivity to Cisplatin by Targeting Survivin Directly and Indirectly Through CUG-BP1. Oncogene, 35(16), 2087-2097.
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4

Apoptosis Induction by miR-214-3p and Cisplatin

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After transfection with pre-miR-214-3p (12 nM) for 48 hours, TE7 cells were treated with Cisplatin (15 μM) (clinical grade) for 24 hours and stained with fluorescein-labeled Annexin-V (FITC) and Propidium Iodide (PI) (TACS Annexin V FITC Apoptosis Detection kit, Trevigen Inc, Gaithersburg, MD). Briefly, cells were washed twice with cold PBS and spun at 300g for 5 min at RT. 106 cells were resuspended in 100μl Annexin V incubation reagent (10 μl 10× binding buffer, 2 μl PI, 1 μl Annexin V FITC and 87 μl deionized water per sample) and incubated in the dark for 15 minutes at RT. At the end of incubation, 400 μl of 1× binding buffer was added (per 100 μl reaction). Samples were then analyzed for FITC Annexin-V positive, PI negative or FITC Annexin-V positive, PI positive by flow cytometry (BD Accuri C6). For caspase activation assays, cells were exposed to Cisplatin following transfection with pre-miR-214-3p for 24 hours and cleaved caspase was measured by Western blot using rabbit anti-caspase-3 (Cell Signaling, Danvers, MA). Caspase enzyme activity was measured in triplicate for each experiment, using instant ELISA kit (eBioscience Inc., San Diego, CA), following the manufacturer's instructions.
Phatak P., Byrnes K.A., Mansour D., Liu L., Cao S., Li R., Rao J.N., Turner D.J., Wang J.Y, & Donahue J.M. (2015). Overexpression of miR-214-3p in Esophageal Squamous Cancer Cells Enhances Sensitivity to Cisplatin by Targeting Survivin Directly and Indirectly Through CUG-BP1. Oncogene, 35(16), 2087-2097.
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5

Serum CRP and Plasma IL-6 Assessment

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Serum high sensitivity CRP concentrations were analyzed using a commercially available Instant ELISA kit (eBioscience), according to the manufacturer's instructions. The limit of detection was 3 pg/ml. The intra-and inter-assay coefficients of variability for serum CRP measurements were 6% and 8%, respectively. Plasma IL-6 concentrations were analyzed using a commercially available high sensitivity ELISA kit (eBioscience), according to the manufacturer's instructions. The limit of detection was 0.03 pg/ml. The intra-and inter-assay coefficients of variability for plasma IL-6 measurements were 8% and 7%, respectively.
Dittrich K., Boedeker K., Kluczniok D., Hindi Attar C., Winter S.M., Roepke S., Heim C., Herpertz S.C, & Bermpohl F. (2021). Elevated inflammatory markers in women with remitted major depressive disorder and the role of early life maltreatment. Brain, behavior, and immunity, 97.
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6

Plasma MCP-1 Quantification

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For determination of MCP-1 concentration in plasma (pg/ml), Instant ELISA kit from eBioscience® was used. Assay procedures were applied as described in the product manual. Rat chemokine present in the samples binds to anti-rat chemokine antibodies adsorbed to the microwells. The reaction of secondary biotin-conjugated anti-rat chemokine antibody was evaluated by streptavidin-HRP. Tetramethyl-benzidine reaction with HRP bound to immune complex was measured at 490 nm in comparison with reference wavelength 620 nm (microplate reader MRX II, Dynex, USA). The results were calculated from standard calibration curve on internal standards.
Gardi C., Bauerova K., Stringa B., Kuncirova V., Slovak L., Ponist S., Drafi F., Bezakova L., Tedesco I., Acquaviva A., Bilotto S, & Russo G.L. (2015). Quercetin reduced inflammation and increased antioxidant defense in rat adjuvant arthritis. Archives of biochemistry and biophysics, 583.
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7

Biomarkers of Metabolic and Inflammatory Status

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Plasma fetuin A levels were measured by ELISA kit (Biovendor, Brno, Czech Republic) with sensitivity 0.104 ng/ml. Plasma fetuin B levels were measured by ELISA kit (Biovendor, Brno, Czech Republic). Sensitivity was 0.19 ng/ml. Plasma FGF21 levels were measured by ELISA kit (Biovendor, Brno, Czech Republic). Sensitivity was 7 ng/ml. C-reactive protein (CRP) levels were measured by instant ELISA kit (eBioscience, Vienna, Austria) with sensitivity 3 pg/ml. The intra-and interassay variabilities for all assays were between 5.0 and 10.0 %.
Serum levels of cytokines were measured by multiplex assay MILLIPLEX MAP Human High Sensitivity T Cell Panel (Merck KGaA, Darmstadt, Germany). Sensitivity for IFN-γ was 0.48 pg/ml, IL-10 was 0.56 pg/ml, IL-6 was 0.11 pg/ml, IL-8 was 0.13 pg/ml and for TNF-α was 0.16 pg/ml. The intra-and interassay variabilities for all assays were between 5.0 and 15.0 %.
Biochemical parameters (urea, creatinine, uric acid, total bilirubin, ALT, AST, ALP, GGT, glycated hemoglobin -HbA 1c , HDL cholesterol, total cholesterol, triglycerides) were measured and LDL cholesterol was calculated at the Department of Biochemistry, General University Hospital, Prague, Czech Republic by standard laboratory methods.
Šimják P., Cinkajzlová A., Anderlová K., Kloučková J., Kratochvílová H., Lacinová Z., Kaválková P., Krejčí H., Mráz M., Pařízek A., Kršek M, & Haluzík M. (2018). Changes in plasma concentrations and mRNA expression of hepatokines fetuin A, fetuin B and FGF21 in physiological pregnancy and gestational diabetes mellitus. Physiological research, 67(Suppl 3).
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8

Tunable Secretion of Super-IL2 with STS-Notch

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Example 25

This Example describes experiments performed to demonstrate tunable secretion of super-IL2 with STS-variants of Hinge-Notch.

Primary human T-cells were activated with anti-CD3/anti-CD28 Dynabeads (Gibco) and transduced with a lentiviral construct Hinge-Notch receptor with inducible super-IL2 under Gal4-UAS control (FIG. 18A). Hinge-Notch receptors containing 3 different STS variants (NRG1, Notch1, Notch2) were tested against a no HingeNotch control. T cells were co-incubated with MCAM+ CD19+ K562 cells in media lacking IL-2, and at the indicated timepoints, supernatant IL-2 was measured using the Instant ELISA Kit (Invitrogen) according to manufacturer's protocols with a microplate reader (Tecan). Red dotted line indicates a standard concentration of IL-2 used for culturing T cells. Graded secretion of super-IL2 was achieved by activation of STS-tuned HingeNotch receptors.

For FIG. 18B, primary human T-cells were generated with an additional lentiviral vector expressing a CAR against MCAM. Enhanced uptake of IL-2 by CAR-expressing cells resulted in loss of supernatant IL2 in CAR-only and NRG1-STS Hinge-Notch T cells. In contrast, greater induction of super-IL2 by Notch1 and Notch2-STS based receptors initially outpaces this uptake, before proliferation and K562 elimination reduces supernatant levels.

US11202801B2. Notch receptors with hinge domain (2021-12-21). The Regents of the University of California [US]. Inventors: Kole T. Roybal [US], Raymond Liu [US], Iowis Zhu [US].
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9

Cytokine Quantification by ELISA

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TNF-α and IP-10 were quantified in cell supernatants using DuoSet ELISA kits from R&D Systems and human IL-1β using Instant ELISA™ Kit from Invitrogen, according to the manufacturer’s instructions. Absorbance was read with a BioTek Synergy HT Microplate Readers.
Pizzuto M., Lonez C., Baroja-Mazo A., Martínez-Banaclocha H., Tourlomousis P., Gangloff M., Pelegrin P., Ruysschaert J.M., Gay N.J, & Bryant C.E. (2019). Saturation of acyl chains converts cardiolipin from an antagonist to an activator of Toll-like receptor-4. Cellular and Molecular Life Sciences, 76(18), 3667-3678.
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10

Tunable Secretion of Super-IL2 via Hinge-Notch

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Example 25

This Example describes experiments performed to demonstrate tunable secretion of super-IL2 with STS-variants of Hinge-Notch.

Primary human T-cells were activated with anti-CD3/anti-CD28 Dynabeads (Gibco) and transduced with a lentiviral construct Hinge-Notch receptor with inducible super-IL2 under Gal4-UAS control (FIG. 18A). Hinge-Notch receptors containing 3 different STS variants (NRG1, Notch1, Notch2) were tested against a no HingeNotch control. T cells were co-incubated with MCAM+ CD19+ K562 cells in media lacking IL-2, and at the indicated timepoints, supernatant IL-2 was measured using the Instant ELISA Kit (Invitrogen) according to manufacturer's protocols with a microplate reader (Tecan). Red dotted line indicates a standard concentration of IL-2 used for culturing T cells. Graded secretion of super-IL2 was achieved by activation of STS-tuned HingeNotch receptors.

For FIG. 18B, primary human T-cells were generated with an additional lentiviral vector expressing a CAR against MCAM. Enhanced uptake of IL-2 by CAR-expressing cells resulted in loss of supernatant IL2 in CAR-only and NRG1-STS Hinge-Notch T cells. In contrast, greater induction of super-IL2 by Notch1 and Notch2-STS based receptors initially outpaces this uptake, before proliferation and K562 elimination reduces supernatant levels.

US11617766B2. Notch receptors with hinge domain (2023-04-04). The Regents of the University of California [US]. Inventors: Kole T. Roybal [US], Raymond Liu [US], Iowis Zhu [US].
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