Hesperidin was procured from Sigma Aldrich, USA. ELISA kits for IL-6, IL-1β, and TNF-α were purchased from Lifespan Biosciences, Inc., WA, USA. iNOS, NO, SOD, and MDA kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, Jiangsu, China).
Tnf α
Manufactured by LifeSpan BioSciences
Sourced in United States
TNF-α is a protein that plays a key role in the inflammatory response. It is a cytokine secreted by various cells, including macrophages, lymphocytes, and endothelial cells. TNF-α is involved in the regulation of immune cells and the induction of inflammation.
Automatically generated - may contain errors
Lab products found in correlation
Mda kit, by Nanjing Jiancheng (1 mentions)
Hesperidin, by Merck Group (1 mentions)
Il 18, by Thermo Fisher Scientific (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Il 1β, by BioLegend (1 mentions)
Il 10, by LifeSpan BioSciences (1 mentions)
Bca kit, by Thermo Fisher Scientific (1 mentions)
Ecl western blotting analysis system, by Merck Group (1 mentions)
Bcl 2, by LifeSpan BioSciences (1 mentions)
Il 1β, by GeneTex (1 mentions)
Bcl2 associated x (bax), by Enzo Life Sciences (1 mentions)
Ripa reagent, by Solarbio (1 mentions)
β actin, by GeneTex (1 mentions)
Interleukin 6 (il 6), by GeneTex (1 mentions)
P iκbα, by GeneTex (1 mentions)
4 protocols using tnf α
1
Hesperidin Biomarker Evaluation Protocol
2
Quantification of Inflammatory Cytokines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell culture medium supernatants from each group were collected by the end of treatment, and IL-1β, TNF-α, FasL and MMP-7 protein levels in supernatants were measured using monkey IL-1β, TNF-α, FasL and MMP-7 ELISA kits (all from LifeSpan BioSciences, Seattle, WA, United States) according to the manufacturer’s instructions. After adding stop solution to terminate reaction, the absorbance of substrate was read by a spectrophotometer at a wavelength of 450 nm immediately. Each experiment was performed four times in three replicates.
3
Measuring Inflammatory Cytokine Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ELISA kits were used for assessing the levels of the following inflammatory parameters: IL-10 and TNF-α (LifeSpanBioSciences, Inc., Seattle, WA, United States), IL-6 (R&D System, Minneapolis, MN, United States), IL-1β (BioLegend, San Diego, CA, United States), IL-18 (eBioscience, Vienna, Austria), following manufacturers’ instructions. All cytokine assays were determined in duplicate.
4
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RIPA reagent (Solarbio) and BCA kit (#23235, Thermo Fisher Scientific) were chosen to conduct protein extraction and concentration quantification, several times. The different proteins of every sample were separated using gel electrophoresis and then transferred onto polyvinylidene difluoride (PVDF, Bedford, MA, USA) membranes. Then, they were covered and maintained in 5% fat-free milk at room temperature for 2 h. The primary antibodies included Bax (No. ADI-AAM-140-E, Enzo Life Sciences, Inc., Farmingdale, NY, USA), Bcl-2 (No. ADI-AAM-072-E), TNF-α (No. LS-B6067, LifeSpan BioSciences, Seattle, WA, USA), IL-6 (No. LS-C746886), IL-1β (No. 503513, San Diego, CA, USA), p-IκBα (phospho Ser32/Ser36, No. GTX79042, GeneTex, Irvine, CA, USA), t-IκBα (No. GTX27545), p-p65 (phospho Ser468, No. GTX32256), t-p65 (No. GTX107678), and β-actin (No. GTX629630). After incubation with above primary antibodies at 4℃ overnight, secondary antibodies were used to block at room temperature for 1 h. Signals of protein were developed with an ECL™ Western Blotting Analysis system (Sigma). The intensity was analyzed by ImageJ software (version 146; National Institutes of Health, Bethesda, MD, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!