Products

Price Inquiry

Welcome! For price inquiries, please feel free to contact us through the form at the bottom. We will get back to you as soon as possible.

Magic Red Cathepsin B Assay

Product Name:

Price:

Inquiry

Description:

Quantitate and monitor intracellular cathepsin-B activity over time in vitro. The Magic Red substrate in this assay fluoresces red upon cleavage by active cathepsin enzymes. Analyze the fluorescent signal using fluorescence microscopy or a fluorescence plate reader.

Shelf Life:

12 months

Detection Method:

Fluorescence Plate Reader, Fluorescence Microscope

How To Use:

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 (480 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 33342, DAPI, or an antibody.
8. If desired, fix 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.

Introduction:

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 assay kit detects cathepsin B activity.

To use Magic Red, add the substrate directly to the cell culture media, incubate, and analyze. Because it is cell permeant, it easily penetrates the cell membrane and the membranes of the internal cellular organelles – no lysis or permeabilization steps are required. If cathepsin enzymes are active, the Magic Red substrate is cleaved and the cresyl violet fluorophore 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 with a fluorescence plate reader. Hoechst 33342 and Acridine Orange are included in the kit to detect nuclear morphology and lysosomal organelle structure, respectively.

Kit Contents:

25 Tests
• Magic Red Substrate (MR-RR2), 25 Test Vial.
• Acridine Orange, 0.5 mL.
• Kit Manual.

100 Tests
• Magic Red Substrate (MR-RR2), 100 Test Vial.
• Hoechst 33342, 1 mL.
• Acridine Orange, 0.5 mL.
• Kit Manual.

Measurement:

592 nm / 628 nm

Sample:

Cell culture

Storage Conditions:

Store at 2-8°C.
avoid freeze/thaw cycles

Target:

Cathepsin-B

Testing Results:

Cathepsin B-positive THP-1 cells fluoresce red after staining with MR-(RR)2. Red fluorophore concentrates inside lysosomes (Dr. Brian W. Lee, ICT).

It should be noted that our products are for research purposes only. Not suitable for any clinical use.

Online Inquiry