9151
Green Fluorescent Cathepsin B Assay
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£275.00
- SKU:
- 9151
- Anwendung:
- Flow Cytometry
- Physischer Zustand:
- Cell Culture
- Aufreinigung:
- Cathepsin activity in THP-1 monocytes. Intracellular cathepsin activity was detected in THP-1 monocytes that were either untreated to show baseline cathepsin activity (green histogram) or were treated with the cathepsin inhibitor CA-074 Me (blue histogram) for 3 hours to reduce cathepsin activity. After treatment, cell populations were stained for 1 hour at 37°C with R110-(RR)2 . After incubation with the R110-(RR)2 substrate, the samples were analyzed by flow cytometry with an Attune NxT flow cytometer equipped with a blue laser (488 nm excitation and 530/30 emission filter). An overlay of the histograms is shown below. Treatment with CA-074 Me decreased the fluorescence signal detected compared to the untreated cell population. Cathepsin activity in THP-1 macrophages. Intracellular cathepsin activity was detected in THP-1 cells. THP-1 cells were seeded onto chamber slides and were then treated with PMA (5 ng/mL for 48 hours) to become macrophage-like. After 48 hours, media containing PMA was replaced with fresh media, and cells were allowed to recover for 4 days, and then were exposed to the cathepsin inhibitor, CA-074 Me for 3 hours (lower row of images), or were untreated (upper row of images). Following treatment, samples were stained with R110-(RR)2 for 1 hour at 37°C and Hoechst 33342 for 15 minutes at room temperature, and then were imaged. Intracellular localization of the hydrolyzed (fluorescent) R110 product was detected using a Logos iRiS Digital Cell Imaging System equipped with a EGFP (Ex 470/30, Em 530/50) and a DAPI (Ex 375/28, Em 460/50) LED filter cubes at 20X. The images below show untreated cells (upper row of images) and inhibitor-treated cells (lower row of images) stained with R110-(RR)2 (left most images), Hoechst 33342 (middle images), or an overlay of both the EGFP and DAPI channels (rightmost images).
- Lagerbedingungen:
- 2-8[o]C, -20[o]C
- Hersteller:
- ImmunoChemistry Technologies
- Immunogen:
- Cathepsin B
- Weitere Details:
- The quenched substrate Rhodamine 110-(RR)2, fluoresces green upon cleavage by active cathepsins, allowing the monitoring of intracellular cathepsin activity in vitro using live whole cells. Analyze using flow cytometry or fluorescence microscopy., ICT
- Versandbedingungen:
- Blue Ice