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11 protocols using human il 17 elisa kit

1

Quantification of Inflammatory Cytokines by ELISA

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The inflammatory cytokines including tumor necrosis factor alpha (TNF‐α), interleukin‐1 beta (IL‐1β), interleukin‐10 (IL‐10), and interleukin‐17 (IL‐17) in the plasma were determined by Enzyme‐linked immunosorbent assay (ELISA) with ELISA kits, including Human TNF‐α ELISA Kit (Thomas, KHC3014), Human IL‐1β ELISA Kit (Thomas, BMS224HS), Human IL‐10 ELISA Kit (Abcam, ab100549), and Human IL‐17 ELISA Kit (Abcam, ab83707), which were ready‐to‐use, according to the instructions of manufacturer. In brief, firstly, samples or standards were added to the 96‐well plates, followed by the antibody mix. After incubation, the wells were washed to remove unbound material. TMB substrate (tetramethylbenzidine, TMB) was added, generating blue coloration. This reaction was then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal was generated proportionally to the amount of bound analyte (Biotek), and the intensity was measured at 450 nm. Each reaction was run in triplicate by the same operator. Standard curves were prepared before the antibody reaction, and the standard curve range of Human TNF‐α ELISA Kit (Thermo, KHC3014), Human IL‐1β ELISA Kit (Thermo, BMS224HS), Human IL‐10 ELISA Kit (Abcam, ab100549), and Human IL‐17 ELISA Kit (Abcam, ab83707) were 0.5‐32pg/mL, 0.16 ‐10 pg/mL, 2.34‐150 pg/mL, and 3.12‐100 pg/mL, respectively.
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2

Cytokine Profiling Using ELISAs

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The levels of IL-1, IL-6, IL-17 and NF-kB were measured using Human IL-6 ELISA® Kit, Human IL-1 beta ELISA® Kit, Human IL-17 ELISA® Kit, and NFkB p65 Transcription Factor Assay® Kit (Abcam, Cambridge, MA, USA), respectively, according to the manufacturer’s instructions. Moreover, the amount of TGF-β and TNF-α were measured by Human TGF-beta 1 Quantikine ELISA® Kit (Minneapolis, MN, USA) and Human TNF Alpha PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA), respectively, according to the manufacturer’s instructions.
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3

Quantifying Inflammatory Cytokines in Sepsis

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For the sepsis patients, the inflammatory cytokine levels in plasma samples were detected by enzyme‐linked immunosorbent assay (ELISA). The commercial ELISA kits including Human Tumor Necrosis Factor‐α (TNF‐α) ELISA Kit, Human Interleukin (IL)‐1β ELISA Kit, Human IL‐6 ELISA Kit, Human IL‐8 ELISA Kit, Human IL‐10 ELISA Kit, and Human IL‐17 ELISA Kit were purchased from Abcam (Cambridge, USA). The procedures were conducted in accordance with the manufacturers’ instructions.
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4

Inflammatory Cytokine Profiling in Acute Ischemic Stroke

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The inflammatory cytokines including tumor necrosis factor (TNF‐α), interleukin‐6 (IL‐6), and interleukin‐17 (IL‐17) in AIS patients’ serum samples were detected by ELISA, which were carried out referring to the instructions of ELISA Kits. The brief procedures were the same as SIRT2 detection. The ELISA Kits used in the study were as follows: human TNF‐α ELISA Kit (Abcam, Cambridge, USA), human IL‐6 ELISA Kit (Abcam, Cambridge, USA), and human IL‐17 ELISA Kit (Abcam, Cambridge, USA).
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5

Quantifying Inflammatory Cytokine Levels

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The levels of tumor necrosis factor alpha (TNF‐α), interleukin‐1 beta (IL‐1β), and interleukin‐6 (IL‐6), and interleukin‐17 (IL‐17) in the plasma of participants were determined by the enzyme‐linked immunosorbent assay (ELISA) with the use of ELISA kits (Abcam) including Human TNF alpha ELISA Kit, Human IL‐1 beta ELISA Kit, Human IL‐6 ELISA Kit, and Human IL‐17 ELISA Kit. Standard curves were made by using standards provided in the kits, and the cytokine concentrations were appointed from the standard curves by use of linear regression analysis. All experiments were performed according to the protocols provided by the manufacturer.
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6

Measurement of Th-related Cytokines

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Th-related cytokines in peripheral blood and gingival crevicular fluid samples were detected. The gingival crevicular fluid was collected as follows. After the supragingival plaque and dental calculi were removed, aseptic cotton was used to keep the moisture, and the surface was wiped dry using sterile cotton balls. The dental face was blown dry using a pneumatic gun. Then #30 absorbent paper was gently inserted into the periodontal pocket on the side of cheek until there was slight resistance. After 30 s, the paper was taken out and placed into sterile tubes. The same operation was performed at an interval of 30 s. If there was visible blood or saliva contamination, the paper was discarded, and the sample was re-taken after 30 s. Gingival crevicular fluid samples were cryopreserved. Then the levels of IL-2, IL-4, IL-10, IL-17, tumor necrosis factor-β (TNF-β) and IFN-γ were determined using human IL-2 ELISA kit (ab174444), human IL-4 ELISA kit (ab215089), human IL-10 ELISA kit (ab46034), human IL-17 ELISA kit (ab119535), human TNF-β ELISA kit (ab229202) and human IFN-γ ELISA kit (ab46025) (Abcam, USA).
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7

Aspergillus GM and Immune Markers in BALF and Plasma

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plasma samples were collected from each subject before treatment initiation. Bronchoalveolar lavage was performed on the intrapulmonary area according to the results of the chest CT examination to obtain bronchoalveolar lavage fluid. The plasma and BALF samples were assayed for Aspergillus GM antigen by ELISA (Platelia Aspergillus Kit, Bio-Rad Laboratories). ELISA also was used to detect plasma Dectin-1 (Human Dectin-1 ELISA Kit, Thermo Fisher) and IL-17 (Human IL-17 ELISA Kit, Abcam) levels following the standard methods.
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8

APACHE II Scoring in Severe Acute Pancreatitis

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It is well documented that APACHE II scores are associated with the severity of SAP and its complications and mortality.15 (link) The APACHE II scores, measured using a Microsoft® APACHE II graded computer program, were recorded. Venous blood samples were taken after a 12-h fast to measure biochemical parameters. The levels of fasting blood glucose (BG), serum amylase (AMY), albumin (ALB), blood calcium, CRP, tumour necrosis factor (TNF)-α, interleukin (IL)-6, IL-10 and IL-17 were all measured using the following colorimetric and enzyme-linked immunosorbent assay (ELISA) kits from Abcam, Cambridge, UK: Glucose Assay Kit (ab65333), Amylase Assay Kit (ab102523), Human Albumin ELISA Kit (ab179887), Calcium Assay Kit (ab102505), Human C Reactive Protein ELISA Kit (ab99995), Human TNF Alpha ELISA Kit (ab181421), Human IL-6 ELISA Kit (ab178013), Human IL-10 ELISA Kit (ab46034) and Human IL-17 ELISA Kit (ab119535) according to the manufacturer’s instructions. The total white blood cell (WBC) count was measured using an automatic haematology analyser (Sysmex XT-2000i; Sysmex Corporation, Kobe, Japan) according to the manufacturer’s instructions. Observed indices also included the time it took to achieve relief of abdominalgia, time of first defaecation, hospitalization time, complications and the cure rate.
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9

Cytokine Profiling in SLE Patients

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IL-18, IL-6 and IL-17 concentrations were measured in the cell supernatants and serum of patients with SLE by ELISA using Human IL-18 ELISA kit (cat. no. ab215539; Abcam), human IL-6 ELISA kit (cat. no. ab46042; Abcam) and human IL-17 ELISA kit (cat. no. ab119535; Abcam), respectively.
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10

Quantifying IL-17 in Cell Cultures

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The concentration of IL-17 in culture supernatants were measured by Human IL-17 ELISA Kit (ab119535, Abcam) according to the manufacturer’s instructions. Optical density values were read at 450 nm using ELx800 Absorbance Microplate Reader (BioTek, VT, USA).
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