神奇紅組織蛋白酶 L 檢測試劑盒

在體外隨時間推移定量和監測細胞內組織蛋白酶 L 的活性。該試驗中的 Magic Red 試劑在被活性組織蛋白酶切割後發出紅色螢光。使用熒光顯微鏡或螢光盤式分析儀分析螢光訊號。

Background

Elevated cathepsin enzyme activity in serum or the extracellular matrix can signify a number of pathological conditions. ICT’s Magic Red assay kits are used by researchers seeking to quantitate and monitor cathepsin activity in cultured cells and tissues. This Magic Red kit detects cathepsin L The Magic Red reagent MR-FR2 enters each cell in a non-fluorescent state. If cathepsin enzymes are active, the Magic Red substrate is cleaved and the cresyl violet fluorophone will become fluorescent upon excitation. As enzyme activity progresses and more Magic Red substrate is cleaved, the signal will intensify, allowing researchers to watch the color develop over time. Samples can be analyzed by fluorescence microscopy or fluorescent plate reader. Samples may also be analyzed with Acridine Orange or Hoechst stain to detect lysosomal organelle structure or nuclear morphology respectively.

Reagent Name

MR-FR2

Target

Cathepsin L

Excitation / Emission

592 nm / 628 nm

Method of AnalysisF

luorescence Plate Reader, Fluorescence Microscope

Sample Type

Cell culture, tissue

Storage2-8°C

Country of Origin

United States

Protocols

1. Prepare samples and controls

2. Reconstitute Magic Red by adding 50 µL DMSO.

3. Dilute Magic Red 1:10 by adding 450 µL diH2O.

4. Add 20 µL Magic Red to each sample (~ 500 µL aliquot of cultured cells).

5. Incubate while protected from light.

6. Watch color start to develop within 15 minutes.

7. If desired, label with additional stains, such as Hoechst, DAPI, Acridine orange, or an antibody.

8. If desired, fix or embed cells.

9. Analyze with a fluorescence microscope or fluorescence plate reader. Magic Red has an optimal excitation at 592 nm and emission at 628 nm.

****If working with adherent cells, please see the manual for additional protocols.

Kit Contents

Kit# 941: 25 TestsMagic Red Reagent (MR-FR2), 1 25-test vial, #6137 Hoechst 33342 Stain, 1 mL, #639 Acridine Orange Stain, 0.5 mL, #6130 Kit Manual