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HIF1a Reporter Lentivirus
$595.00
Fluorescent HRE reporter lentivirus (HIF1a): High-quality, lentiviral hypoxia-response element (HRE) reporter system that provides a sensitive fluorescent readout for human/mouse hypoxia-inducible factor 1 alpha (HIF-1 alpha, HIF1A, or HIF-1a) activity in transduced cells. The transcription factor (TF) reporter construct is preferentially activated by HIF-1a (vs HIF-2a) and may be useful for discriminating HIF1a and HIF2a transcriptional activities and for studying hypoxia pathway activation. The lentiviral particles are purified by PEG precipitation and sucrose gradient centrifugation, and are ideal for studying HIF-1a activity in difficult-to-transfect cells including primary and/or thawed cells.
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Email: info@lipexogen.com
Available Options:
Specifications
Key Advantages:
- High Sensitivity – LipExoGen HIF1a Reporter lentiviral particles are made using a novel vector platform based on the third generation system. The transcription factor’s hypoxia response elements (HRE) are arranged as DNA tandem repeats upstream of the minimal TATA promoter-driven reporter, and downstream of an optimized minimal enhancer (pc) of the human CMV promoter. When the signal pathway/TF is activated, the mini enhancer synergizes with TF binding to the response elements (up to 8 repeats in some products, depending on strength of reporter activation) to amplify expression of the fluorescent (GFP/RFP) or luciferase (Luc) reporter, with minimal enhancement of background. As a result, the reporter system provides a highly sensitive readout for signaling pathway or specific transcription factor activation in human and mouse cells.
- Easily Establish Stable Reporter Cell Lines – The reporter lentiviral particles are ultra-purified and concentrated to high-titer by PEG purification and sucrose gradient centrifugation to allow for efficient transduction of difficult-to-transfect cells, including primary and/or freeze-thawed cells. Stable cell lines are easily generated through puromycin or blasticidin selection.
- Discovery Made Easy – Signal pathway or specific transcription factor activity can be detected by fluorescence, making LipExoGen TF Reporter lentiviral particles more practical than traditional luciferase reporters and/or biochemical assays. Pathway/TF activation can be readout directly by fluorescence microscopy in living cell cultures, thus paving the way for unexpected discoveries.
- Readout On Flow – Fluorescent reporter activation can also be readout by flow cytometry, providing more versatility in data acquisition for labs with different instruments.
- Best Value – LipExoGen lentiviral particle products are made using optimized lentiviral vectors developed in-house, which allows us to provide the highest quality products while retaining competitive prices. These high-titer lentiviral particles feature a highly sensitive fluorescent reporter system which has been validated to read out the activity of the indicated transcription factor or signaling pathway.
- Same Cost For Custom Lentivirus – You can easily request any combination of reporter (GFP/RFP/Luc) and selection marker (puromycin/blasticidin) for this product, without additional cost, by contacting us. To view our complete list of vectors, click here.
Product Data:
Figure 1 (thumbnail). HEK293FT cells were co-transfected with HIF1a-TAG-Puro (left panels) or HIF1a-TAR-BSD (right panels) plasmids in combination with plasmid for stable HIF1a (bottom panels) or control (vector, top panels) for 24-36 hours. Fluorescence microscopy images were then taken to asses fluorescent reporter expression in the live cells. Stable means mutant HIF1a to confer resistance to normoxia-induced degradation.
Figure 2. Monitoring HIF1a activity in vivo with luciferase HCT116 cells were transduced with HIF1a-TAL-Puro (LTV-0006-3S) and puromycin selected to establish a stable cell line. The luciferase reporter activity can be detected in vivo using common imaging software. A reduction in reporter signal following administration of an HIF1a transcriptional inhibitor (48 h after treatment) depicts how the product could be used for lead validation or to screen for novel HIF1a inhibitors in vivo.
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Details:
| LTV-0006 | |
| High-titer Lentiviral Particles, third generation, VSV-G pseudotyped | |
| Serum-free RPMI-1640, frozen solution | |
| PEG precipitation and sucrose gradient centrifugation | |
| One vial | |
| 380 μl/vial | |
| 1-5×10^6 TU/vial, depending on the variant | |
| 3×10^8 VP/ml | |
| -80oC | |
| Hypoxia/HIF1α signaling pathway | |
| Species | Human/mouse |
| Tandem repeats of response elements coupled to minimal TATA promoter and upstream proprietary enhancer | |
| Hypoxia response elements (HRE), preference to HIF1α over HIF2α | |
| GFP, RFP, or Firefly Luciferase | |
| Promoter (Driving Selection Marker) | EF1α |
| Puromycin (Puro) or Blasticidin (BSD), or a fluorescent protein for select products | |
| Monitor HIF1α transcriptional activity. The fluorescent reporter enables convenient readout using flow cytometry, fluorescence microscopy, etc. |
*Based on infectivity on HEK293FT cells transduction units (TU).
Related Products
Hypoxia/HIF Pathway:
HIF1a shRNA Lentivirus (Human/mouse)
HIF2a shRNA Lentivirus (Human/mouse)
HIF2a Transcriptional Reporter Lentivirus (Human/mouse)
Hif1a-PPN ORF cDNA Lentivirus (Mouse)
Vector Diagram
Recommended Controls
Reporter Negative Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven reporter. The construct is the same as the TF Reporters except that it lacks the transcriptional response elements which drive signal pathway/TF-specific reporter expression. The reporter negative control lentiviral particles allow to establish a baseline for background reporter activity and determine specificity of any treatments to activate the reporter.
Reporter Positive Control Lentivirus – Ready-to-transduce lentiviral particles with constitutive expression of the reporter. The reporter positive control lentivirus is useful for transduction optimization studies, especially if the cells are very sensitive or difficult-to-transduce.
Renilla Luciferase Internal Control Lentivirus – Ready-to-transduce lentiviral particles expressing minimal TATA box-driven Renilla luciferase (RLuc).
Publications
Custom Orders
If you require a modification to one of our products, such as a change in reporter or other vector component, please contact us. Examples of customization options are shown in the table below. Feel free to request something not in the table.
Additional Custom Service Options
- Send us your cells and we can establish a stable NFAT reporter cell line for you using this product. Learn more.
- ORF cDNA plasmids featured in the product figures are available upon request.
- Ultra-high concentration NFAT reporter virus can be provided upon request in your choice of medium and volume (i.e. for in vivo applications).
Background
Background
HIFs are implicated in diverse cellular processes including oxygen sensing and adaptation, metabolism, Warburg effect, stress response, chemoresistance, immune function, angiogenesis, tumor growth, epithelial-mesenchymal transition (EMT), cancer stem cell maintenance and self-renewal, extracellular matrix (ECM), and metastasis. As a result, the HRE reporter lentivirus can be used as a valuable tool for studying the hypoxia pathway and cellular response to changes in oxygen tension, and other various cellular processes regulated by the HIF pathway. The vector construct has been validated to provide sensitive fluorescent readout in response to HIF1a transcriptional activity (Fig. 1).
Additional Information
Additional Information
| HIF1A | |
| hypoxia inducible factor 1 subunit alpha | |
| n | NM_001530 |
| Homo sapiens/mus musculus | |
| Alias | HIF1A, hypoxia inducible factor 1 subunit alpha, Hypoxia Inducible Factor 1, Alpha Subunit (Basic Helix-Loop-Helix Transcription Factor), Class E Basic Helix-Loop-Helix Protein 78, Basic-Helix-Loop-Helix-PAS Protein MOP1, Hypoxia-Inducible Factor 1-Alpha, PAS Domain-Containing Protein 8, Member Of PAS Protein 1, HIF-1-Alpha, BHLHe78, PASD8, MOP1, Hypoxia-Inducible Factor 1 Alpha Isoform I.3, Hypoxia Inducible Factor 1 Alpha Subunit, Hypoxia-Inducible Factor1alpha, Member Of PAS Superfamily 1, ARNT Interacting Protein, ARNT-Interacting Protein, HIF-1alpha, HIF1-ALPHA, HIF1-Alpha, BHLHE78, HIF-1A, HIF1 |
| Annotation Page | https://www.ncbi.nlm.nih.gov/gene/?term=hif1a |
| Gene IDs | HGNC: 4910, Entrez Gene: 3091, Ensembl: ENSG00000100644, OMIM: 603348, UniProtKB: Q16665, GC14P061695 |
| Entrez Gene Summary | “This gene encodes the alpha subunit of transcription factor hypoxia-inducible factor-1 (HIF-1), which is a heterodimer composed of an alpha and a beta subunit. HIF-1 functions as a master regulator of cellular and systemic homeostatic response to hypoxia by activating transcription of many genes, including those involved in energy metabolism, angiogenesis, apoptosis, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. HIF-1 thus plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Alternatively spliced transcript variants encoding different isoforms have been identified for this gene. [provided by RefSeq, Jul 2011]” |