The oxidative stress markers such as malondialdehyde (MDA) and protein carbonyls (PC) and antioxidants such as catalase (CAT) and reduced glutathione (GSH) were measured from the hippocampal lysates using commercially available kits from Cayman chemicals (Arbor, MI). We also measured the transcription factor nuclear factor erythroid 2-related factor 2 (NRF2) from hippocampal lysates using a kit from Signosis (Santa Clara, CA). We followed the manufacturer's instructions for these assays. The total protein concentration in different tissue lysates was measured using a Pierce BCA reagent kit (Thermo Fisher Scientific, Waltham, MA), and the concentration of the markers was normalized with the total protein in respective tissue lysates.
Pierce bca reagent kit
Manufactured by Thermo Fisher Scientific
The Pierce BCA reagent kit is a protein quantification assay used to determine the total protein concentration in a sample. It relies on the bicinchoninic acid (BCA) method, where proteins reduce Cu2+ to Cu+ in an alkaline environment. The Cu+ ions then chelate with BCA to produce a purple-colored complex, which can be measured spectrophotometrically.
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Lab products found in correlation
2 protocols using pierce bca reagent kit
1
Oxidative Stress Biomarkers in Hippocampus
2
Estradiol Regulates Metastasis Signaling
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Previous studies have shown that 17β-estradiol can regulate metastasis and invasion through a rapid ERα36–dependent membrane–mediated pathway involving PLC and PKC.10 (link),16 (link),19 (link),23 (link),25–27 (link) To investigate if these signaling pathways are involved in the effect of E2 on TNBC osteolysis, we evaluated the activity of both PLC and PKC after treatment with E2 and E2-BSA. MDA-MB-231 cells were cultured to confluence on six–well plates and treated with vehicle, 10-8 M, or 10-7 M E2-BSA for 9 min. Cells were harvested immediately in 300 μL PLC lysis buffer or PKC lysis buffer according to the manufacturer’s protocol and assayed using the Amplex Red Phospholipase C Assay Kit (Thermo Fisher Scientific) or PKC Activity Assay Kit (Abcam), respectively. Cell layer lysates were assayed for total protein content using the Pierce BCA reagent kit (Thermo Fisher Scientific). PLC and PKC activities were normalized to total protein content per minute.
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