Pro-inflammatory cytokines such as IL-6 levels were measured using commercial ELISA Kits (Cat.No.:E-EL-H6156; Human IL-6 ELISA Kit; Elabscience Biotechnology Inc., Houston, TX 77079, USA) with a sensitivity level of 0.94 pg/mL and a detection range of 1.56–100 pg/mL, while TNF-α levels were measured using commercial ELISA kits (Cat.No.: E-EL-H0109; Human TNF-α ELISA Kit; Elabscience Biotechnology Inc., Houston, TX 77079, USA) with a sensitivity level of 4.69 pg/mL and a detection range of 7.81–500 pg/mL. Several studies have validated the accuracy of commercial ELISA kits used to analyze IL-6 and TNF-α levels [22 (link),46 (link),47 (link)].
Human il 6 elisa kit
Manufactured by Elabscience
Sourced in United States
The Human IL-6 ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay designed for the measurement of human interleukin-6 (IL-6) levels in various biological samples, such as cell culture supernatants, serum, and plasma.
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Lab products found in correlation
Human tnf α elisa kit, by Elabscience (4 mentions)
Human il 8 elisa kit, by Elabscience (1 mentions)
Bn 2 nephelometric analyzer, by Siemens (1 mentions)
P iκb, by Cell Signaling Technology (1 mentions)
Lipopolysaccharide lps from e coli 055 b5, by Merck Group (1 mentions)
P jnk jnk, by Cell Signaling Technology (1 mentions)
Human il 8, by R&D Systems (1 mentions)
Inf γ, by R&D Systems (1 mentions)
Neutrophil elastase, by Cell Signaling Technology (1 mentions)
Histone h3, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
Il 1β, by R&D Systems (1 mentions)
Il 10, by R&D Systems (1 mentions)
Human ace2 elisa, by Abcam (1 mentions)
Psicheck 2 vector, by Promega (1 mentions)
M 280 streptavidin coated magnetic beads, by Merck Group (1 mentions)
Varioskan lux microplate reader, by Thermo Fisher Scientific (1 mentions)
Dual luciferase reporter assay kit, by Promega (1 mentions)
Rneasy minelute cleanup kit, by Qiagen (1 mentions)
8 protocols using human il 6 elisa kit
1
Quantifying Pro-inflammatory Cytokines Using ELISA
2
Plasma IL-6 Quantification in ICU Patients
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Venous blood samples for the determination of plasma levels of IL-6 were obtained were obtained at admission and at 48 h after admission in the ICU.
The blood samples (100 µL) were collected in tubes that contained EDTA (10 mmol/L) to prevent activation of the system of coagulation. The tubes were centrifuged immediately for 10 min at 3,000 rpm, and the plasma was stored in aliquots at –70 °C.
The measurement of IL-6 was performed in duplicate in the blood samples by using an enzyme immunoassay with Human Il-6 ELISA kit (Elabscience Biotechnology Inc. USA). The detection range was 7.81–500 pg/mL, with intra and interassay coefficients of variation below 5%. Readings were carried out in an automated optical Spectramax reader at 450 nm. The sensitivity of the assay was <2 pg/mL. The serum levels of this cytokine are below the detection limit in healthy subjects. Results were related to a dose-response curve obtained with recombinant human IL-6 and expressed as picograms per milliliter, normal values of IL-6 corresponding to<9.7 pg/mL [23 (link)].
The blood samples (100 µL) were collected in tubes that contained EDTA (10 mmol/L) to prevent activation of the system of coagulation. The tubes were centrifuged immediately for 10 min at 3,000 rpm, and the plasma was stored in aliquots at –70 °C.
The measurement of IL-6 was performed in duplicate in the blood samples by using an enzyme immunoassay with Human Il-6 ELISA kit (Elabscience Biotechnology Inc. USA). The detection range was 7.81–500 pg/mL, with intra and interassay coefficients of variation below 5%. Readings were carried out in an automated optical Spectramax reader at 450 nm. The sensitivity of the assay was <2 pg/mL. The serum levels of this cytokine are below the detection limit in healthy subjects. Results were related to a dose-response curve obtained with recombinant human IL-6 and expressed as picograms per milliliter, normal values of IL-6 corresponding to<9.7 pg/mL [23 (link)].
3
Cytokine Secretion Profiling of Stimulated Lymphocytes
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The serum of all subjects was collected, sealed and stored in a -80 °C refrigerator for later detection. The lymphocyte culture supernatants stimulated by lipopolysaccharide for 72 h, lipopolysaccharide and CR for 72 h were collected, sealed and stored in a -80 °C refrigerator for later detection. The secretion levels of IL-6, IL-8, IL-17 cytokines in the samples were determined using the relevant ELISA kits according to the manufacturer's instructions(HumanIL-17A ELISA Kit, Human IL-6 ELISA Kit, Human IL-8 ELISA Kit, All purchased from Elabscience, 96 T).
4
Measuring Inflammatory Markers in Blood Samples
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For objective data, 2 mL of the patients’ venous blood was collected before the ITM surgery. ESR values were measured using the Vision ESR analyzer (YHLO Biotech Co., Shenzhen, China), and CRP values were measured using the BN II nephelometric analyzer (Siemens Healthcare Diagnostics, Marburg, Germany). IL-6 levels (pg/mL) were determined using the Human IL-6 Elisa Kit (Elabscience Biotechnology Co., Wuhan, China), and TNF-a levels (pg/mL) were determined using the Human TNF-α Elisa Kit (Elabscience Biotechnology Co., Wuhan, China).
5
Dehydrozingerone Attenuates Inflammatory Response
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Dehydrozingerone (> 99% purity) bought from TCI chemicals and structure of the compound was shown in Fig. S1A, Lipopolysaccharide (LPS from E. Coli 055: B5) purchased from Sigma Aldrich (USA). Dexamethasone was purchased from Sigma-Aldrich, USA. C-DNA synthesis kit, SYBR green mix purchased from Takara bioscience INDIA. Antibodies GAPDH, JNK P-JNK, c-JUN, p-NF-κB, NF-κB, p38, pP38, Histone-H3, IκB, p-IκB, MPO, Neutrophil Elastase were purchased from Cell signalling technology (USA), ELISA kits (IL-6, CCL2, IL-10, INF-γ, Human-IL-8 and IL-1β were bought from R&D systems. Human IL-6 ELISA kit was procured from Elabsciences (Houston, USA). Secondary Antibodies were procured from Jackson Immuno research private limited (USA). TUNEL kit was procured from Merk Millipore.
6
Quantifying ACE2 and IL-6 in S1 Spike Exposure
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The levels of ACE2 and IL-6 were measured using ELISA after 24 hours of S1 spike protein exposure. The level of ACE2 was measured using Abcam Human ACE2 ELISA following the manufacturer protocol (Abcam, Cat. Ab235649) while Human IL-6 ELISA kit (Elabscience Biotechnology, E-EL-H0102) was used to measure the levels of IL-16. The levels of ACE2 and IL-6 were measured in triplicates.
7
Exploring VSMC Cytokine Release and Regulation
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The releases of IL-6 and TNF-α from human VSMCs were detected via enzyme-linked immunosorbent assay (ELISA) using human IL-6 ELISA Kit (#E-EL-H0102c, Elabscience, Wuhan, China) or human TNF-α ELISA Kit (#E-EL-H0109c, Elabscience) according to the manufacturer's protocols. Dual-luciferase reporter assays: Sequences of circ_ 0044073-wild type (WT), circ_0044073-mutation (MUT), AURKA-3'UTR-WT, and AURKA-3'UTR-MUT were ligated into empty psiCHECK-2 vectors (Promega, Madison, WI, USA). Human VSMCs were transfected with a luciferase reporter together with miR-377-3p mimic or miR-NC. The lysates of the transfected human VSMCs were analyzed using a Varioskan LUX microplate reader (Thermo Fisher) and a dual-luciferase reporter assay kit (Promega). RNA pull-down assays: Biotinylated miR-377-3p and miR-NC (80 nM) were transfected into human VSMCs for 48 hours. Then, the lysates of human VSMCs were incubated with M-280 streptavidin-coated magnetic beads (Sigma-Aldrich) at 4°C for 3 hours. The bound RNAs were then extracted using an RNeasy MinElute Cleanup Kit (Qiagen, Valencia, CA, USA) and subjected to qRT-PCR analysis.
8
Cytokine Detection in Cell Culture
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Enzyme-linked immunosorbent assay (ELISA) was performed to detect pro-inflammatory cytokines in cell culture supernatants with the recommended protocol. The following ELISA kits were used: Human IL-6 ELISA Kit (Elabscience, E-EL-H6156), Human TNF-α ELISA Kit (Elabscience, E-EL-H0109c).
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