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Cat B 750 FAST |
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Agent Description
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11098 |
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Cat B 750 FAST including 1 vial, 10 doses (based on a 20-25 g mouse)- Fluorescence agent specifically activated by cathepsin B proteinases (Cat B). Optically silent in the unactivated state, becoming highly fluorescent when activated. |
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Cat B 750 FASTTM is a member of a family of activatable fluorescent imaging agents comprising a novel architecture, termed F.A.S.T. (Fluorescent Activatable Sensor Technology) that confers an improved pharmacokinetic profile with a broader range of early imaging timepoints. This architecture also offers higher target specific signal with reduced background.
Cat B 750 FAST is a Cathepsin B activatable agent that is optically silent upon injection and produces fluorescent signal after cleavage by Cathepsin B produced by inflammatory cells and tumor cells. Cat B 750 FAST may be used to monitor inflammation, tumor activity, progression of disease and the efficacy of therapeutic treatment in applications such as oncology, inflammation, cardiovascular disease and some neurological diseases.
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Enzyme Panel Cat B 750 FAST
Cat B 750 FAST: 0.5 µM final concentration of agent in the optimized buffer for each enzyme and 0.05 µM final concentration of activated enzyme. |
Macrophages Activate Agent: FACS + Inhibitors
Raw 264.7 mouse macrophages were incubated with Cat B 750 FAST (1 μM final concentration). Cathepsin inhibitors were added 1 hour before addition of the agent: E64d cell-permeable cysteine/calpain inhibitor and CA-074Me specific cell-permeable cathepsin B inhibitor. Cells were analyzed by fluorescence microscopy
Cat B 750 FAST is activated by RAW macrophages. The pan-cathepsin inhibitor E64d and the specific cathepsin B inhibitor CA-074Me significantly reduce the activation.
Pharmacokinetic Profile of Cat B 750 FAST
Cat B 750 FAST in plasma shows a calculated t(1)1/2 = 2-4 min.
Cat B 750 FAST Representative Images (4 nmoles)-Kinetics
Subcutaneous tumor model; Same mice are shown at 6h and 24h.
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Imaging intestinal adenomas in APCM in mice
The C57BL/6J-ApcMin/J strain is highly susceptible to spontaneous intestinal adenoma formation similar to patients with familial adenomatous polyposis. C57BL/6J-ApcMin heterozygous mice raised on a high fat diet develop in excess of 30 adenomas throughout the intestinal tract and most die by 120 days of age. ApcMin (18 weeks-old,10 weeks on high fat diet) and control C57BL/6 mice were injected i.v. with 5 nmoles Cat B 750 FAST. The GI tract was excised 6h later and imaged on the FMT2500 in reflectance mode. Significantly higher Cat B 750 FAST signal in the intestine of APCMin mice, associated with the presence of adenomas. |
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Imaging atherosclerosis in ApoE-/- mice
Apolipoprotein E (ApoE) deficient mice were generated by targeted inactivation of the apoE gene. These mice develop atherosclerotic lesions (particularly after being fed a high fat diet) resembling those seen in humans. Eight week-old female ApoE-/- mice fed a high fat diet for 20 weeks or C57BL/6 age-matched wild type (WT) controls were injected i.v. with 8 nmoles Cat B 750 FAST. Aortas were excised 6 hrs later and imaged on the FMT2500 in reflectance mode. Significantly higher Cat B 750 FAST signal in aortas of ApoE-/- mice, associated with the presence of atherosclerotic plaques (mainly in the aortic root and arch). |
Cat B 750 FAST Properties & Specifications
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Cat B 750 FAST Characteristics
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1. Absorbance and fluorescence maxima of activated 2. As determined by RP-HPLC and measuring absorbance |
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Normalized absorbance (grey), excitation (blue) and fluorescence emission (red) spectra of Cat B 750 FAST in 1x PBS. |
Material (Needs to be reconstituted)
Cat B 750 FAST: Each vial contains 48 nmol of Cat B 750 FAST in dry solid form. Cat B 750 FAST has been filtered through a 0.2 µm filter prior to drying. Reconstitute Cat B 750 FAST with 1.2 mL of 1 x PBS before injecting into animals. The packaged material provides sufficient reagent for imaging approximately 10 mice (weighing ~25 grams each) when using the recommended dose of 4 nmols (100 µL) of Cat B 750 FAST per mouse. Some applications like Atherosclerosis imaging require a higher dose.
Properties
The physical characteristics of Cat B 750 FAST can be found in table above.
Storage and Handling
Upon receipt, Cat B 750 FAST should be STORED AT 2-8 oC AND PROTECTED FROM LIGHT.
When stored and handled properly, Cat B 750 FAST is stable for up to 3 months in dry solid form.
Before opening the vial check to ensure that all of the solid material is at the bottom of the vial.
After reconstituting with PBS, gently swirl the solution to ensure that the solid is fully in solution.
Once reconstituted with 1 x PBS, the solution is stable up to 10 days when stored at 2-8 ºC and protected from light.
In Vivo Imaging and Applications
Cathepsin B is a cysteine protease involved in the degradation of the extracellular matrix and may be one of the key factors in cancer progression. Cathepsin B correlates with invasiveness and metastatic capabilities in many tumors. In breast cancer specifically, high expression levels of cathepsin B have been linked to highly aggressive tumors and poor clinical outcome. Cathepsin B is also upregulated in a variety of inflammatory cells (including eosinophils, neutrophils, and macrophages), and may provide an inflammation readout that indicates the potential vulnerability of Atherosclerotic plaques in Cardiovascular disease.
The generally recommended procedure for in vivo imaging with Cat B 750 FAST is administration via intravenous injection and imaging 6 hours post injection. Earlier and later timepoints may be appropriate for some disease models, and the optimal imaging timepoint for any application should be determined empirically. Some applications like imaging of atherosclerotic plaques require an 8 nmol dose and may benefit from a later imaging timepoint.
Cat B 750 FAST enables imaging of Cathepsin B activity in applications including:
Cardiovascular disease,
Oncology,
Inflammation,
Certain neurological diseases
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Part #
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Agent Description
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11098 |
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Cat B 750 FAST including 1 vial, 10 doses (based on a 20-25 g mouse)- Fluorescence agent specifically activated by cathepsin B proteinases (Cat B). Optically silent in the unactivated state, becoming highly fluorescent when activated. |
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