Cat.No. : AD00362Z
Titer: ≥1x10^10 IFU/mL / ≥1x10^11 IFU/mL / ≥1x10^11 VP/mL / ≥1x10^12 VP/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | AD00362Z |
| Description | Human Adenovirus Type5 (dE1/E3) expressing Nanog Homeobox under CMV promoter. No fusion tag, pre-made adenovirus, ready to ship and ready to use format. |
| Target Gene | NANOG |
| Product Type | Adenoviral particle |
| Insert | NANOG |
| Titer | Varies lot by lot, for example, ≥1x10^10 IFU/mL, ≥1x10^11 IFU/mL, ≥1x10^11 VP/mL etc. |
| Size | Varies lot by lot, for example, 250 ul, 500 ul, 1 mL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality adenovirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between adenovirus particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in adenovirus production, especially for applications in animal studies and gene therapy. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced adenovirus particles to ensure regulatory compliance. |
| Sterility | Creative Biogene ensures that adenovirus products are free of any bacterial, fungal and other microbial contamination. |
| Ad5 E1 Detection | All Creative Biogene adenoviruses are PCR tested to ensure that there are no detectable E1 sequences in the particles, which could be from revertants or external E1 contamination. |
| RCA Assays | Adenovirus products originating at Creative Biogene are guaranteed to have undetectable replication-competent adenovirus (RCA). This quality control measure is important because there is always the possibility of wild-type contamination due to revertants or environmental sources. |
| PFU Titering | All purified adenovirus preparations are tested for infectious titer. Creative Biogene's PFU test takes a few days longer but counts true plaques in HEK cells rather than estimating PFU titers via IHC staining or TCI50 of infected cells. |
Here, the researchers explored the expression of nanoprotein homeobox (NANOG) in thoracic aortic dissection (TAD) and its role in regulating the phenotypic switch of human aortic vascular smooth muscle cells (VSMCs). The experimental results showed that NANOG and OPN were highly expressed in the aortic wall and VSMCs in TAD, and both were accompanied by the phenotypic switch of VSMCs. Overexpression of NANOG induced upregulation of matrix metalloproteinase 2, a synthetic marker of VSMCs, and downregulation of contractile markers α-smooth muscle actin and smooth muscle 22α. Overexpression of NANOG also enhanced the proliferation, migration and anti-apoptosis ability of VSMCs. The results also showed that these functions of NANOG were achieved through OPN, and NANOG directly upregulated OPN by binding to its promoter region.
Smooth muscle cells (VSMCs) were isolated from control samples and transfected with NANOG overexpression adenovirus (Ad-NANOG) to investigate the effects of NANOG on osteopontin (OPN) expression and smooth muscle cell phenotype. Transfection effects were detected by fluorescence observation, qRT-PCR, and Western blot (Figure 1A, B, and D). The expression level of NANOG protein relative to β-actin was measured, and the results were similar to those in TAD VSMCs. The researchers first evaluated the expression changes of related genes at the mRNA level compared with VSMCs transfected with Ad-GFP. After 48 hours of transfection, the mRNA expression level of OPN in Ad-NANOG VSMCs increased to 2.30 ± 0.20 (Figure 1C). In Ad-NANOG VSMCs, the VSMC synthesis marker MMP2 increased to 11.17±2.98, but the VSMC contraction markers α-SMA and SM22α decreased to 0.71±0.13 and 0.41±0.12, respectively (Figure 1F-H). Western blot results confirmed these changes at the protein level (Figure 1E, I-K).
Figure 1. Overexpression of NANOG up-regulated the expression of OPN and VSMCs synthetic MMP2, whereas the expression of VSMCs contractile markers α-SMA and SM22α were down-regulated. (An Z, et al., 2017)
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The NANOG adenovirus helped us achieve robust reprogramming in our stem cell models. The purity and activity were excellent—definitely a top choice for pluripotency research!
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