ELISA was performed to detect mIL-1β (BGK10749, Peprotech, Cranbury, NJ, USA), mIL-6 (BGK08505, Peprotech), and mTNF-α (BGK06804, Peprotech) in RAW 264.7 cell culture supernatant, following the provided manufacturer’s instructions. The cell culture supernatant was diluted with PBS (10, 50, 100, and 200 times) to confirm cytokine expression levels. According to the test results, the cell supernatant was diluted and loaded onto each well coated with the antibody in advance, and then incubated at 37 °C for 1 h 30 min. Next, 1× biotinylated antibody and 1× avidin-biotin-peroxidase complex were added sequentially. Afterward, the color-developing reagent was loaded and incubated for 10–30 min, and the stop solution was added to terminate the reaction. Finally, a Bio-RAD x-MarkTM spectrophotometer (Table 1 ; Bio-Rad Laboratories, USA) was used to measure the pro-inflammatory cytokine levels.
Bgk06804
Manufactured by Thermo Fisher Scientific
Sourced in United States
The BGK06804 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed for use in scientific research and analysis applications. The core function of this product is to provide a controlled environment for various experimental and testing procedures. No further details about the intended use or specific features of this product are available.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using bgk06804
1
Quantifying Cytokine Levels in RAW 264.7 Cells
2
RNA Isolation and Cytokine Analysis
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Total RNA was isolated from RAW264.7 cells and NHDFs were treated with PBS or EVs using Labozol reagent (Cosmo Gentech, Seoul, Korea) according to the manufacturer's protocol. The purified RNA was quantified using a NanoPhotometer (Implen, Munich, Germany). cDNA synthesis was performed with 2μg of total extracted RNA using an M‐MuLV Reverse Transcription Kit (Cosmo Gentech, Seoul, Korea). Quantitative real‐time PCR was performed using EzAMP PCR 2x Master Mix (ELPIS Biotech, Daejeon, Korea) according to the manufacturer's recommendations. The culture medium of treated RAW264.7 cells was analysed for tumour necrosis factor (TNF)‐α (Peprotech, BGK06804, Cranbury, NJ, USA) and Interleukin (IL)‐6 (Peprotech, BGK08505) production using ELISA. A Bio‐RAD x‐Mark spectrophotometer was used to measure the absorbance at 450 nm.
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