Infected [EPEC (WT), HS4, and EPEC mutants] and uninfected (control) Caco-2 cells were incubated with labeled (0.1 μCi) or labeled plus unlabeled (1mM AA) AA containing Krebs-Ringer (KR) buffer (in mM: 1.23 MgSO4:7H2O, 133 NaCl, 10 HEPES, 4.93 KCl, 5 glucose, 0.85 CaCl2 dihydrate, 5 glutamine, 10 MES hydrate; pH 7.4) for 30 min in a 37°C water bath. To determine AA uptake in jejunum and colon tissues, mice were euthanized to extract the jejunum and colonic sheets, which were then opened longitudinally and immediately incubated in KR buffer with either labeled (0.1 μCi) or labeled plus unlabeled (1mM AA) AA at 37°C for 7 min as described [35 (link)]. Cells or tissues were lysed with 1N NaOH and heat treated at 80°C for 15 min, followed by neutralization with 10N hydrochloric acid. Subsequently, radioactive content was determined (liquid scintillation counter; Beckman Coulter, Brea, CA). Protein concentrations of cells or intestinal tissues were determined with Protein Assay kit reagents (Bio-Rad, Hercules, CA).
Protein assay kit reagents
Manufactured by Bio-Rad
The Protein Assay kit reagents are a solution-based system designed for the quantification of protein samples. The reagents enable the colorimetric detection and measurement of protein concentrations. The core function of this product is to provide a reliable and accurate method for determining the protein content in various samples.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using protein assay kit reagents
1
Quantifying Arachidonic Acid Uptake
2
Protein Separation and Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein separation was performed using the iNtRON Biotechnology kit. First, 10 or 20 µM tianeptine was added to SH-SY5Y cells. After 6, 24, or 72 hours of culture, cells were washed with PBS and harvested by 2000-3000 rpm centrifugation. To 5×106 SH-SY5Y cells, 400 µL Pro-Prep was added with mixing and cells were incubated on ice for 20 minutes. After 13,000 rpm centrifugation for 5 minutes at 4℃ , proteins in the upper layer were collected and stored at -20℃ for Western blotting and quantitative analysis. Protein assay kit reagents (Bio-Rad) were diluted 1:5 with distilled water, and 1, 0.5, 0.1, 0.01, or 0 mg/mL BSA standards were set up. After placing 10 µL of BSA standards in 96-well plates, 200 µL working solution was added. Prepared BSA standards and protein samples to be measured were placed at room temperature for 5 minutes. Absorbance was measured with an ELISA reader at 595 nm, and protein concentration was measured using the standard curve.
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!