Human tnf alpha elisa kit

Plasma cytokine concentrations were measured using the Human TNF alpha ELISA kit (ab181421; Abcam, Cambridge, UK) and the Human IL-10 ELISA Kit (ab185986; Abcam, Cambridge, UK). ELISA assays were performed in accordance with the manufacturer’s instructions. All samples were tested in duplicate, and results were expressed as pg/mL.

The levels of IL-4 and TNF-alpha were measured using the Human IL-4 ELISA Kit or the Human TNF-alpha ELISA Kit, respectively, according to the manuals provided (Abcam, Cambridge, UK).

Human IL-6 ELISA Kit (ab178013), Interferon-gamma ELISA Kit (ab100538), Human TNF-alpha ELISA Kit (ab181421), and Human IL-8 ELISA Kit (ab108869) were purchased from Abcam (Cambridge, UK). Cytokine levels were determined according to the manufacturer’s protocol. Briefly, 50 µL of each sample and blank was placed in 96-well plates and incubated at room temperature for 1 h. Then, each well was washed 3 times with wash buffer and 50 μL of 1× biotinylated antibody for 30 min. TMB development solution was, then, added and incubated for 10 min. Then, 100 μL of stop solution was added, and absorbance was measured at 450 nm using an Infinite M200 Microplate Reader (Tecan Infinite M200 Pro; Tecan GmbH, Männedorf, Switzerland).

Measurement of TNF-α levels in serum was performed using the Human TNF alpha ELISA Kit (catalog no. ab46087) from Abcam (Discovery Drive, Cambridge Biomedical Campus, Cambridge, CB2 0AX, UK), according to the manufactures’ instructions. Briefly, blood samples were centrifuged at 2000× g for 10 min and the collected serum was stored at −20 °C until further analysis.

Determination of the content of pro-inflammatory cytokines TNFα, IL1β, and endogenous β-amyloid peptide Aβ40 in the lysate of human peripheral blood mononuclear cells was performed after 1 and 24 h of incubation with rAβ42 by immunoenzymatic method with spectrophotometric detection on a FL600 microplate multimodal reader (BioTek) using test kits “Human TNF alpha ELISA Kit” (Abcam, cat. no. ab181421), “Human IL-1 beta ELISA Kit” (Abcam, cat. no. ab214025) and “Amyloid beta 40 Human ELISA Kit” (Thermo Scientific, cat. No. #KHB3481) respectively. NP-40 buffer (Thermo Scientific, cat. no. J60766.AP) was used for cell lysis. Determination of total protein in cell lysate was carried out by the direct UV spectrophotometric method on a scanning spectrophotometer Ultrospec 3,100 pro (Biochrom). The content of cytokines and endogenous Aβ40 was calculated in ng/g of total protein.

After treatments, the protein expressions of IL‐6 or TNF‐α in the cell culture supernatants were detected by the Mouse IL‐6 enzyme‐linked immunosorbent assay (ELISA) Kit (Abcam, USA), the Human IL‐6 ELISA Kit (Abcam, USA), the Mouse TNF alpha ELISA Kit (Abcam, USA), or the Human TNF alpha ELISA Kit (Abcam, USA) according to the manufacturer's instruction.

ELISAs were performed to detect the levels of TNF-α secreted into supernatants from the control and Iso2-overexpression liver cancer cells. Briefly, the supernatants were collected and centrifuged at 2000× g for 5 min to clear the cells for further study. The levels of TNF-α were comparatively analyzed using a Human TNF alpha ELISA Kit (Abcam, Waltham, MA, USA) following the manufacturers’ instructions.