Cat.No. : AD00372Z
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. | AD00372Z |
| Description | Human Adenovirus Type5 (dE1/E3) expressing High-mobility Group Box 1 with C-terminus 6xHN tag under a CMV promoter. C-terminus 6xHN tag, pre-made adenovirus, ready to ship and ready to use format. |
| Target Gene | POU4F3 |
| Product Type | Adenoviral particle |
| Insert | POU4F3, C-fusion with 6xHN tag |
| 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. |
| Gene Name | High-mobility Group Box 1 |
| Gene Symbol | HMGB1 |
| Gene ID | 3146 |
| mRNA Refseq | NM_002128 |
Lipopolysaccharide (LPS) is known to mediate angiogenic effects in endothelial cells. However, the underlying mechanisms remain unclear. The studies here show that LPS can induce the secretion of high-mobility group protein box 1 (HMGB1) from human pulmonary microvascular endothelial cells (HPMECs). Knockdown and overexpression of HMGB1 by adenoviral vectors effectively inhibited and promoted LPS-induced HPMEC migration and capillary-like tube formation, respectively. On the other hand, HMGB1 had an inhibitory effect on LPS-inhibited platelet-endothelial cell adhesion molecule (CD31) and p120 catenin (p120) expressions; knockdown of HMGB1 reversed this effect. These results suggest that LPS and HMGB1 have a functional synergistic effect in angiogenesis. Mechanistically, the physical interaction of LPS with HMGB1 mediated the dissociation of p120, β-catenin, and γ-catenin from vascular endothelial cadherin (VE-cadherin) without affecting VE-cadherin expression. The synergistic effect of LPS and HMGB1 was closely related to the ERK/P38/Src signaling pathway, as reflected by the reduced migration and capillary-like tube formation in HPMECs treated with signaling pathway inhibitors. Together, these findings reveal a novel mechanism by which LPS and HMGB1 synergistically regulate the angiogenic behavior of endothelial cells.
To detect the release of HMGB1 in HPMECs regulated by LPS, the researchers tested adenoviral vectors expressing HMGB1 (Ad-HMGB1) and shHMGB1 (Ad-shHMGB1) in HPMECs. As shown in Figures 1A-D, these adenoviral vectors effectively regulated the expression of HMGB1, indicating that they can be used for gain-of-function and loss-of-function assays. Next, the researchers sought to explore the effects of HMGB1 on LPS-mediated epithelial cell migration and capillary-like tubule formation, two processes that are key to the initiation of angiogenesis. Wound healing assays showed that knockdown of HMGB1 by Ad-shHMGB1 significantly reduced LPS-induced HPMEC migration compared with the Ad-control group (Figures 1E and F), while overexpression of HMGB1 by Ad-HMGB1 led to a significant increase in the extent of HPMEC migration, regardless of the presence or absence of LPS (Figures 1E and F). Likewise, Ad-HMGB1 and Ad-shHMGB1 significantly increased and decreased the number of capillary-like microvascular structures in the absence or presence of LPS stimulation, respectively (Figures 1 G and H). These data indicate that HMGB1 is essential and sufficient to regulate the angiogenic behavior of LPS-stimulated endothelial cells, suggesting that LPS and HMGB1 have a functional synergistic role in angiogenesis.
Figure 1. Individual and combined effects of LPS and HMGB1 on HPMECs migration and capillary-like tube formation. (Liu Z, et al., 2017)
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The HMGB1 adenovirus induced strong inflammatory signaling in our immune assays. The viral quality was top-notch, and delivery was efficient. Perfect for studying sepsis or autoimmune diseases.
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