rabbit anti human cyp3a4  (Proteintech)

Journal: European journal of clinical pharmacology

Article Title: A cocktail probe approach to evaluate the effect of hormones on the expression and activity of CYP enzymes in human hepatocytes with conditions simulating late stage of pregnancy.

doi: 10.1007/s00228-023-03489-1

Figure Lengend Snippet: Fig. 1 CYP3A4 activity in freshly isolated human hepatocyte cultures treated with Vehicle (0.1% DMSO), third trimester plasma (LH) and predicted liver concentrations of female hormones (HH), Rifampin (Rif, 10 µM) and Ketoconazole (Ket, 10 µM). Fold induction values are expressed relative 6β- hydroxylation of testosterone formation rates compared to vehicle treated hepatocytes in the respective batch of hepatocytes. HU1552, HU1527, HU1593, HU1632 and HU12-010 represent CYP3A4 activity in individual batch of hepatocytes and Mean represents the average of activity in 5 batches of hepatocytes. Treatments were compared using ANOVA followed by Tukeys' multi- ple comparison test; ****p < 0.0001

Article Snippet: Rabbit anti-human primary monoclonal antibodies against CYP3A4 and β-actin and anti-rabbit IgG linked with horseradish peroxidase were obtained from Cell Signaling Technology, Inc. (Danvers, MA).

Techniques: Activity Assay, Isolation, Clinical Proteomics, Comparison

Journal: European journal of clinical pharmacology

Article Title: A cocktail probe approach to evaluate the effect of hormones on the expression and activity of CYP enzymes in human hepatocytes with conditions simulating late stage of pregnancy.

doi: 10.1007/s00228-023-03489-1

Figure Lengend Snippet: Fig. 5 mRNA levels of CYP3A4 measured by qRT-PCR and normal- ized to mRNA expression of GAPDH. Corrected mRNA expression values are expressed as fold increase over vehicle treatment. CYP3A4 mRNA expression in hepatocytes from four donors (HU 1522, HU1527, HU1593 and HU1632) treated with 0.1% DMSO (Vehicle), third trimester plasma (LH) and predicted liver (HH) concentrations of female hormones and collected after 72 hr treatment (with media change every 24 hours). HU1552, HU1527, HU1593 and HU1632 represent mRNA levels in individual batch of hepatocytes and Mean represents the average of mRNA levels in 4 batches of hepatocytes. Treatments were compared using ANOVA followed by Tukeys' multi- ple comparison test; *p < 0.05

Article Snippet: Rabbit anti-human primary monoclonal antibodies against CYP3A4 and β-actin and anti-rabbit IgG linked with horseradish peroxidase were obtained from Cell Signaling Technology, Inc. (Danvers, MA).

Techniques: Quantitative RT-PCR, Expressing, Clinical Proteomics, Comparison

Journal: European journal of clinical pharmacology

Article Title: A cocktail probe approach to evaluate the effect of hormones on the expression and activity of CYP enzymes in human hepatocytes with conditions simulating late stage of pregnancy.

doi: 10.1007/s00228-023-03489-1

Figure Lengend Snippet: Fig. 6 Spearman correlation of CYP3A4 protein and mRNA expres- sion in freshly isolated human hepatocytes treated with 0.1% DMSO (Vehicle), third trimester plasma (LH) and predicted liver (HH) con- centrations of female hormones (with media change every 24 hours). CYP3A4 protein levels were determined by Western blot followed by densitometric analysis and CYP3A4 mRNA expression was estimated using qRT-PCR. Symbols and line represent individual treatment and regression line respectively

Article Snippet: Rabbit anti-human primary monoclonal antibodies against CYP3A4 and β-actin and anti-rabbit IgG linked with horseradish peroxidase were obtained from Cell Signaling Technology, Inc. (Danvers, MA).

Techniques: Isolation, Clinical Proteomics, Western Blot, Expressing, Quantitative RT-PCR

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: Biocompatibility of 5% composite SFC scaffolds (w/v). The viability staining and H&E staining of cells in both scaffolds (A) . SEM images (B) showing the cell growth pattern on the surface of both scaffolds. IHC detection (C) of ALB and CYP3A4 expression of C3A cells in both scaffolds. Quantitative analysis of IHC staining (D) and functional gene expression of C3A cells in both scaffolds (E) . (* p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP 2 (dilution, 1:50; Proteintech), and Alexa Fluor ® 647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Staining, Expressing, Immunohistochemistry, Functional Assay, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: Functional gene and protein expression in different culture conditions using RT-qPCR (A) and IF staining (B) . Higher functional gene expression was found in the 3D co-cultures than in the 2D cultures and 3D monocultures (A) . iHepLPC-Heps in both groups expressed ALB, CYP3A4, and MRP 2 after 14 days of culture (B) . (* p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP 2 (dilution, 1:50; Proteintech), and Alexa Fluor ® 647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Functional Assay, Expressing, Quantitative RT-PCR, Staining, Gene Expression

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: Functional evaluation of iHepLPC-Heps on composite SFC scaffolds. Albumin secretion of different hepatic cultures was assayed using ELISA (A) . Urea synthesis of iHepLPC-Heps cultured under different conditions (B) . Induction of CYP3A4 (C1) and CYP1A2 (C2) expression in response to stimulation with omeprazole and rifampicin, assayed using RT-qPCR. DiI-LDL uptake and CDCFDA staining (D) of iHepLPC-Heps were examined in both groups. (* p < 0.05).

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP 2 (dilution, 1:50; Proteintech), and Alexa Fluor ® 647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Functional Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Expressing, Quantitative RT-PCR, Staining

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Development of Biomimetic Hepatic Lobule-Like Constructs on Silk-Collagen Composite Scaffolds for Liver Tissue Engineering

doi: 10.3389/fbioe.2022.940634

Figure Lengend Snippet: IHC detection of ALB, CYP3A4, and MRP 2 expression in both groups after transplantation. Protein expressions increased gradually over time. Compared with monocultures, the co-culture group showed upregulated expressions at each time point. Scale bar, 50 µm.

Article Snippet: Subsequently, they were incubated overnight at 4°C using the following primary antibodies: mouse anti-human ALB (dilution, 1:50; Santa Cruz Biotechnology, Santa Cruz, CA, United States), rabbit anti-human CYP3A4 (dilution, 1:100; Proteintech, China), rabbit anti-human MRP 2 (dilution, 1:50; Proteintech), and Alexa Fluor ® 647 Mouse anti-Human CD31 (dilution, 1:100; BD Biosciences).

Techniques: Expressing, Transplantation Assay, Co-Culture Assay