{"title":"SignaGen Adenoviral Vectors","description":"\u003cp\u003eReady-to-use recombinant adenoviral vectors from SignaGen Laboratories for transient, high-level gene expression.\u003c\/p\u003e","products":[{"product_id":"ad-cmv-mito-mkeima-bhv22000115","title":"Ad-CMV-mito-mKeima","description":"\u003cstyle\u003e\n.bhp-desc{font-size:16px;color:#1a1a1a;line-height:1.7}\n.bhp-desc h2{font-size:18px;font-weight:700;color:#003366;margin:24px 0 10px;padding-bottom:6px;border-bottom:2px solid #003366}\n.bhp-desc p{margin:0 0 12px}\n.bhp-desc .bhp-spec-table{width:100%;border-collapse:collapse;margin:12px 0 18px}\n.bhp-desc .bhp-spec-table th{width:36%;text-align:left;padding:8px 12px;background:#e8f0fb;color:#003366;font-weight:600;vertical-align:top;border:1px solid #ccd9f0}\n.bhp-desc .bhp-spec-table td{padding:8px 12px;vertical-align:top;border:1px solid #ccd9f0}\n.bhp-desc .bhp-warning-box{background:#fff8e6;border-left:4px solid #e6a817;padding:10px 14px;margin:10px 0 14px;border-radius:0 4px 4px 0}\n.bhp-desc .bhp-cat-pill{display:inline-block;background:#003366;color:#fff;border-radius:20px;padding:3px 12px;font-size:13px;margin-bottom:10px}\n\u003c\/style\u003e\u003cdiv class=\"bhp-desc\"\u003e\n\u003cspan class=\"bhp-cat-pill\"\u003eAdenoviral Vector\u003c\/span\u003e\u003ch2\u003eVector Overview\u003c\/h2\u003e\n\u003cp\u003eAd-CMV-mito-mKeima is a pre-made, replication-incompetent recombinant adenoviral vector that expresses mitochondria-targeted monomeric Keima (mito-mKeima) under the control of the cytomegalovirus (CMV) immediate-early promoter. The reporter is directed to the mitochondrial matrix and reports the delivery of mitochondria to lysosomes during mitophagy. Adenoviral delivery transduces a broad range of dividing and non-dividing mammalian cells and gives high-level, transient expression from a genome that remains episomal rather than integrating into the host chromosome. Transduction efficiency and expression level depend on the target cell type and should be confirmed in each model. The vector is supplied ready to use, removing the need for cloning, packaging, amplification and titration.\u003c\/p\u003e\n\u003ch2\u003eHow mito-mKeima Reports Mitophagy\u003c\/h2\u003e\n\u003cp\u003eKeima is a coral-derived, acid-stable fluorescent protein with a large Stokes shift. Its excitation spectrum depends on pH: in the mildly alkaline mitochondrial matrix (about pH 8.0) excitation near 440 nm predominates, whereas after mitophagic delivery to the acidic lysosome (about pH 4.5) the predominant excitation shifts to about 586 nm. Emission remains at approximately 620 nm in both states. The ratio of the signals collected at the two excitation wavelengths therefore provides a ratiometric measure of the proportion of mitochondria that reside in acidic compartments.\u003c\/p\u003e\n\u003cp\u003eBecause Keima resists lysosomal proteases, the acidic signal accumulates over time and can be read as cumulative mitophagic flux at a single time point. The large Stokes shift also allows mito-mKeima to be imaged together with green-emitting reporters such as GFP-tagged proteins.\u003c\/p\u003e\n\u003ch2\u003eTechnical Details\u003c\/h2\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eCatalog No.\u003c\/th\u003e\n\u003ctd\u003eSL101757\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eVector Backbone\u003c\/th\u003e\n\u003ctd\u003eHuman adenovirus type 5, E1\/E3-deleted (ΔE1\/ΔE3); replication-incompetent\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003ePromoter\u003c\/th\u003e\n\u003ctd\u003eCMV\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eReporter\u003c\/th\u003e\n\u003ctd\u003emito-mKeima (mKeima targeted to the mitochondrial matrix)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSpectral Properties\u003c\/th\u003e\n\u003ctd\u003eExcitation ≈440 nm (neutral\/alkaline pH) shifting to ≈586 nm (acidic pH); emission ≈620 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eExpression Regulation\u003c\/th\u003e\n\u003ctd\u003eConstitutive\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eTiter\u003c\/th\u003e\n\u003ctd\u003e\u0026gt;1E+10 PFU\/mL (lot range 1E+10 to 1E+11 PFU\/mL)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eBuffer\u003c\/th\u003e\n\u003ctd\u003eSignaGen buffer A195\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eBiosafety Level\u003c\/th\u003e\n\u003ctd\u003eBSL-2\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSizes\u003c\/th\u003e\n\u003ctd\u003e50 µL; 200 µL\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003ch2\u003eApplications\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003e\n\u003cstrong\u003eLive Cell Imaging\u003c\/strong\u003e: ratiometric confocal or widefield imaging of mitophagy in living cells, including time-lapse studies of mitochondrial turnover.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eFlow Cytometry\u003c\/strong\u003e: high-throughput quantification of the acidic-to-neutral mito-mKeima signal ratio on instruments configured for dual excitation.\u003c\/li\u003e\n\u003cli\u003e\n\u003cstrong\u003eMitophagy and mitochondrial quality control studies\u003c\/strong\u003e: measuring how genetic perturbations, cellular stress or pharmacological compounds change mitochondrial delivery to lysosomes.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eInterpretation Notes\u003c\/h2\u003e\n\u003cp\u003eThe readout depends on compartmental pH. Treatments that raise lysosomal pH (for example V-ATPase inhibitors such as bafilomycin A1) suppress the acidic signal independently of mitophagy, so include appropriate controls when testing compounds that may affect lysosomal acidification. Image samples live: fixation dissipates the pH gradient on which the readout depends.\u003c\/p\u003e\n\u003ch2\u003eKey References\u003c\/h2\u003e\n\u003cul\u003e\n\u003cli\u003eKatayama H, Kogure T, Mizushima N, Yoshimori T, Miyawaki A. A sensitive and quantitative technique for detecting autophagic events based on lysosomal delivery. \u003cem\u003eChem Biol\u003c\/em\u003e. 2011;18(8):1042-1052. \u003ca href=\"https:\/\/doi.org\/10.1016\/j.chembiol.2011.05.013\" target=\"_blank\" rel=\"noopener\"\u003edoi:10.1016\/j.chembiol.2011.05.013\u003c\/a\u003e\n\u003c\/li\u003e\n\u003cli\u003eSun N, Malide D, Liu J, Rovira II, Combs CA, Finkel T. A fluorescence-based imaging method to measure in vitro and in vivo mitophagy using mt-Keima. \u003cem\u003eNat Protoc\u003c\/em\u003e. 2017;12(8):1576-1587. \u003ca href=\"https:\/\/doi.org\/10.1038\/nprot.2017.060\" target=\"_blank\" rel=\"noopener\"\u003edoi:10.1038\/nprot.2017.060\u003c\/a\u003e\n\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eBiosafety\u003c\/h2\u003e\n\u003cp\u003eAd-CMV-mito-mKeima is a recombinant adenoviral vector handled at BSL-2. Work in a certified biosafety cabinet, wear appropriate personal protective equipment, decontaminate all surfaces and waste, and follow your institutional biosafety committee requirements for viral vector work.\u003c\/p\u003e\n\u003ch2\u003eSafety \u0026amp; Handling\u003c\/h2\u003e\n\u003cdiv class=\"bhp-warning-box\"\u003e\u003cp\u003eStore at −80°C; avoid repeated freeze-thaw cycles. Aliquot on first thaw. For research use only; not for diagnostic or therapeutic use.\u003c\/p\u003e\u003c\/div\u003e\n\u003c\/div\u003e","brand":"SignaGen Laboratories","offers":[{"title":"50 uL","offer_id":67711823118701,"sku":"SL101757-50UL","price":169.0,"currency_code":"USD","in_stock":true},{"title":"200 uL","offer_id":67711823151469,"sku":"SL101757-200UL","price":618.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/Ad-CMV-mito-mKeima-vector-map.png?v=1792258412"}],"url":"https:\/\/www.ebiohippo.com\/collections\/signagen-adenoviral-vectors.oembed","provider":"BioHippo","version":"1.0","type":"link"}