Human tnf α elisa kit

The measurement of TNF-α, IL-6, and IL-8 expression was performed using reagents provided in the Human TNF-α ELISA Kit (ab181421), Human IL-6 ELISA Kit (ab178013), and Human IL-8 ELISA Kit (ab214030) (Abcam, Cambridge, MA, USA), respectively. Furthermore, 2 × 10⁵/well keratinocytes were seeded in 24-well plate with or without 50 μg/mL of kinkéliba leaf extract treatment 24 h before use, with adherent cells collected by scrape after UVA irradiation and the subsequent treatment by kinkéliba leaf extract. The subsequent measurements were conducted according to the manufacturer’s instruction.

Concentrations of serum and HepG2 cells treated with serum drugs of IL1β, IL6, and TNFα were measured by ELISA kit (Abcam, United States, Mouse IL-6 ELISA Kit, ab222503), (Abcam, United States, Human IL-6 ELISA Kit, ab178013), (Abcam, United States, Mouse IL-1β ELISA Kit, ab197742), (Abcam, United States, Human IL-1β ELISA Kit, ab214025), (Abcam, United States, Human TNFα ELISA Kit, ab181421), (Abcam, United States, Mouse TNFα ELISA Kit, ab208348). The serum and cell samples were subjected to the 96-well plates and coupled with antibody cocktails after various treatments and co-incubated for 2 h. After another wash, the TMB Development Solution was incubated for 10 minis. The stop solution was added and the amount was measured by their corresponding calibration curves.

Peripheral venous blood samples were collected at admission, 3 days and 7 days. Serum levels of CRP, IL-1β, IL-6 and TNF-α was detected by ELISA using commercially kits: Human IL-1β ELISA Kit (ab46052), Human IL-6 ELISA Kit (ab229434), Human TNF-α ELISA Kit (ab229399) (all purchased from Abcam, Cambridge, MA, USA) and human CRP ELISA Kit (#MBS3800117, MyBioSource, Inc., San Diego, CA, USA).

Human IL-6 ELISA kit (ab178013, Abcam), human TNF-α ELISA kit (ab181421, Abcam), and human IL-1β ELISA kit (ab229384, Abcam) were used for the detection of these factors in culture medium of BEAS-2B cells. In addition, mouse IL-1β ELISA kit (ab197742, Abcam), mouse IL-6 ELISA kit (ab222503, Abcam), and mouse TNF-α ELISA kit (ab208348, Abcam) were applied for the detection of the expression of these factors in BALF and lung tissues of mice. In addition, mouse FTL ELISA kit (ab157713, Abcam) and mouse FTH1 ELISA kit (ab65080, Abcam) were applied for the detection of the expression of these proteins in BALF and lung tissues of mice. The operation was following the instruction.

Endotoxin content in SWCNT and GO dispersions was assessed using the chromogenic limulus amoebocyte lysate (LAL) assay (Charles River Endosafe, Charleston, SC). The endotoxin content was found to be below FDA-mandated limits of acceptance (0.5 EU/mL) (data not shown). To verify these results, SWCNT samples were also assessed using the TNF-α expression test (TET) that enables unequivocal detection of endotoxin with a sensitivity that is comparable to the conventional LAL assay, but without any interference with the assay, as described previously^{18 (link)}. In brief, HMDM were exposed to nanomaterials or lipopolysaccharide (LPS) in the presence or absence of the specific LPS inhibitor, polymyxin B (10 µM) and TNF-α secretion was measured at 24 h of exposure using a Human TNF-α ELISA Kit purchased from Abcam (Sweden).

The levels of IL-6 and TNF-α in the culture medium of MPC5 cells were determined using human IL-6 ELISA kit (Abcam, ab178013) and human TNF-α ELISA kit (Abcam, ab181421) according to the manufacturer's protocol.