Thermo Scientific DyLight 690 and DyLight 747 In Vivo Apoptosis Detection Kits offer an accurate method to detect apoptosis in small-animal imaging.
These kits contain a DyLight 690- or DyLight 747-conjugated in vivo polycaspase probe, which is a cell-permeable fluorescent detector of active caspases indicative of apoptosis. The probes are comprised of a polycaspase peptide sequence (VAD) and a fluoromethyl ketone (FMK) moiety, which facilitate irreversible binding of the probe to active caspases, and either DyLight 690 or DyLight 747 Dye for near-infrared detection with a range of in vivo instrumentation. These probes are valuable for in vivo drug determination studies, because changes in apoptosis from drug reactivity can be measured by fluorescence imaging in live animals. An increase in apoptosis produces a greater attachment of probes and, subsequently, an increase in fluorescence output. The in vivo technique is nondestructive, thus eliminating the need to remove cells and tissues from sacrificed animals prior to analysis (which induces further apoptosis). Additionally, apoptosis can be monitored in a single animal at multiple time points, reducing the number of animals required for a given study.
Highlights:
- Nontoxic – not harmful to valuable research animals
- Cell-permeable – readily diffuses through all cells as it circulates throughout the body
- Selective for active caspases – eliminates high background and false-positive results common with other caspase detection methods
Applications:
- Small-animal in vivo apoptosis imaging
Product Details:
For optimal results, use DyLight 690 In Vivo Control Reagent (Product No. 84889) or DyLight 747 In Vivo Control Reagent (Product No. 84896) to assess the binding specificity of the polycaspase probe for the apoptotic cells or tissue within the animal. Two control groups are recommended: one group of untreated animals that are injected with the polycaspase probe; and one group of treated animals that are injected with the representative DyLight In Vivo Control Reagent.
Properties of the Thermo Scientific DyLight Polycaspase Probes
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Fluorescent Dye
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Ex/Em†
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円
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MW†††
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DyLight 690
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690/709
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141K
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950
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DyLight 747
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747/776
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223K
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1028
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† Excitation and emission maxima in nanometers (±4nm)
†† Molar extinction coefficient (M-1 cm-1) at the absorption maximum
††† Molecular weight (g/mol) of the probe
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Thermo Scientific DyLight 690 and 747 Apoptosis Detection Kits offer non-invasive active caspase detection. Adult wild-type Balb/c mice were mock-infected or inoculated intranasally with Herpes simplex virus-1 (HSV1), which is known to induce apoptosis in the brain. Mice were then injected intravenously with DyLight 690 (top panel) or 747 (bottom panel) In Vivo Polycaspase Probe 7 days post-infection. In the second experiment, DyLight 747 In Vivo Control Reagent was injected into an HSV1-infected mouse as a reagent control. After 4 or 7 hours (top and bottom panels, respectively), all animals were imaged using a Carestream* In Vivo FX PRO* imager. Strong caspase activity was detected in the brains of animals treated with HSV-1 and injected with either DyLight 690 or 747 caspase probe but not in mock-infected or reagent control animals. Minimal signal was detected in the liver region of the HSV-treated animal injected with either caspase probe, indicating minimal clearance of the probe. Strong signal intensity in the tail indicates where reagent is likely to pool after IV injection.
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