Hsp70 elisa kit

Levels of Hsp70 in the serum and hippocampus were determined by enzyme‐linked immunosorbent assay (ELISA). The Hsp70 ELISA Kit (ab133060, Abcam) was used for assessing Hsp70 levels in the hippocampus. The Hsp70 High Sensitivity ELISA Kit (ab133061, Abcam) was used for assessing Hsp70 levels in mouse serum. The following kits were used for the detection of inflammatory cytokines in the hippocampus: Mouse IL‐6 ELISA kit (ab100713, Abcam) for IL‐6, Mouse IL‐1 beta ELISA Kit (ab197742) for IL‐1ß, and Mouse monocyte chemoattractant protein‐1 (MCP1) ELISA Kit (ab100722) for MCP1. ELISA was performed according to the manufacturer's instruction.

Human IL-18 ELISA Kit (ab215539), Mouse IL-18 ELISA Kit (ab216165), Human IL-1β ELISA Kit (ab214025), Mouse IL-1β ELISA Kit (ab197742), Mouse S100A8 ELISA Kit (ab263886), Mouse S100A9 ELISA Kit (ab213887) and HSP70 ELISA Kit (ab133060) were purchased from Abcam, UK. Human S100A8 (E-EL-H1289c), Human S100A9 (E-EL-H1290c), HSP90 ELISA Kit (E-EL-H1864c), Human HMGB-1 ELISA Kit (E-EL-H1554c), Mouse HMGB-1 ELISA Kit (E-EL-M0676c) were obtained from Elabscience Biotechnology Co., Ltd, China. ATP Assay Kit (S0027) was purchased from Beyotime, China.
Briefly, differentiated U937 or RAW264.7 was stimulated with LPS for 6 h. Next, TDCA was added to the medium at the final concentration of 200 μM for 24 h. Subsequently, the supernatant was collected to measure the level of DAMPs including HSP70, HSP90, S100A8, S100A9, and ATP. All assays were performed following the instructions of the producer.

The SH-SY5Y and MCF-7 cells were seeded in 96-well microplates and treated with 1 μM (c1) and 3 μM (c2) of Au NPs@polyphenols for 24 h and 48 h. After these time points, some samples were used to induce thermal stress by the procedure previously described (ph. 2.8). Then, the cells were lysed, and the HSP-70 ELISA kit (Abcam, ab133060; Abcam, Cambridge, MA, USA) was used, following the manufacturer’s instructions, to build a calibration curve. The samples were read at 450 nm, which is proportional to the amount of analytes bound.