Cat.No. : AD00336Z
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. | AD00336Z |
| Description | Human adenovirus type5 (dE1/E3) expressing FLAG tag under CMV promoter. Used as control, pre-made adenovirus, ready to ship and ready to use format. |
| Target Gene | FLAG |
| Product Type | Adenoviral particle |
| Insert | FLAG |
| 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. |
Primary open-angle glaucoma (POAG) is the second leading cause of irreversible blindness worldwide. Increased endothelin-1 (ET-1) is observed in the aqueous humor (AH) of POAG patients, resulting in increased outflow resistance of the AH. However, the underlying mechanisms remain elusive. Using established in vivo and in vitro POAG models, researchers demonstrate that the water channel aquaporin 1 (AQP1) is downregulated in trabecular meshwork (TM) cells upon exposure to ET-1, which results in a series of glaucomatous changes including actin fiber reorganization, collagen production, extracellular matrix deposition, and altered TM cell contractility. Ectopic expression of AQP1 can reverse ET-1-induced TM tissue remodeling, which requires the presence of β-catenin. More importantly, ET-1-induced AQP1 repression is mediated by the unfolded protein response transcription factor ATF4, which binds to the AQP1 promoter and negatively regulates AQP1 transcription. Therefore, these studies reveal a novel function of ATF4 in controlling the ET-1-induced TM remodeling process in POAG by inhibiting AQP1 transcription.
Here, researchers determined whether AQP1 reduction is responsible for ET-1-induced pathological changes in POAG. Knockdown of AQP1 in HTMCs with si-AQP1 resulted in collapse of actin arcs and formation of thick actin bundles (Figure 1A), altered intracellular and extracellular collagen (Figure 1B-E), and enhanced cellular contractility (Figure 1F), which mimicked the effects of ET-1 on HTMCs. Furthermore, ectopic expression of AQP1 by transfecting HTMCs with an adenoviral vector encoding full-length AQP1 significantly reversed ET-1-induced stress fiber reorganization (Figure 1A), collagen deposition (Figure 1B-E), and enhanced contractility (Figure 1F) in HTMCs compared with vector control (Ad-Flag). Thus, these results suggest that a decrease in AQP1 is required for ET-1-induced TM tissue remodelling in POAG.
Figure 1. AQP1 inhibits ET-1-induced F-actin reorganization, elevated contractility and collagen deposition in HTMCs. (Zhao Y, et al., 2020)
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The FLAG-tagged adenovirus worked flawlessly in our protein interaction studies. Easy to use, and the provided documentation was very helpful. Will purchase again!
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