Testing Data
(Cathepsin B staining in Jurkat T cells after treatment with inhibitors.Jurkat cells were treated with 10 ?M CA-074Me for 3 hours (blue histogram), 10?M Leupeptin for 1 hour (red histogram) or were untreated (green histogram). They were then stained for 1 hour with R110-(RR)2. After incubation with the R110-(RR)2 substrate, the samples were analyzed by flow cytometry using a ZE5 Cell Analyzer (488 nm excitation and a 525/35 emission filter). Treatment with CA-074Me or Leupeptin decreased the fluorescence signal detected compared to the untreated control (see table), which displayed a baseline level of cathepsin activity)
Cathepsin B Detection Kit | Catb-2744 detection kit
Green Cathepsin B Kit
Applications
Flow Cytometry, Functional Assay, Immunofluorescence
Synonyms
Cathepsin B; Green Cathepsin B Kit; Catb-2744 detection kit
Form/Format
Rhodamine 110-(RR)2
Sequence Length
323
Applicable Applications for Catb-2744 detection kit
Flow Cytometry (FC/FACS), Immunofluorescence (IF)
Preparation and Storage
MULTIPLE STORAGE CONDITIONS APPLY ON ARRIVAL.
Store the unopened kit (and each unopened component) according to the storage instructions on each component label.
Store the Rhodamine110-(RR)2 substrate at -20 degree C.
Once reconstituted in DMSO, use the Rhodamine110-(RR)2 substrate immediately or aliquot and store at -20 degree C for 6 months, protected from light.
Avoid repeated freezing and thawing.
Store the unopened kit (and each unopened component) according to the storage instructions on each component label.
Store the Rhodamine110-(RR)2 substrate at -20 degree C.
Once reconstituted in DMSO, use the Rhodamine110-(RR)2 substrate immediately or aliquot and store at -20 degree C for 6 months, protected from light.
Avoid repeated freezing and thawing.
Testing Data
(Cathepsin B staining in Jurkat T cells after treatment with inhibitors.Jurkat cells were treated with 10 ?M CA-074Me for 3 hours (blue histogram), 10?M Leupeptin for 1 hour (red histogram) or were untreated (green histogram). They were then stained for 1 hour with R110-(RR)2. After incubation with the R110-(RR)2 substrate, the samples were analyzed by flow cytometry using a ZE5 Cell Analyzer (488 nm excitation and a 525/35 emission filter). Treatment with CA-074Me or Leupeptin decreased the fluorescence signal detected compared to the untreated control (see table), which displayed a baseline level of cathepsin activity)
Testing Data
(Cathepsin B staining in Jurkat T cells after treatment with inhibitors.Jurkat cells were treated with 10 ?M CA-074Me for 3 hours (blue histogram), 10?M Leupeptin for 1 hour (red histogram) or were untreated (green histogram). They were then stained for 1 hour with R110-(RR)2. After incubation with the R110-(RR)2 substrate, the samples were analyzed by flow cytometry using a ZE5 Cell Analyzer (488 nm excitation and a 525/35 emission filter). Treatment with CA-074Me or Leupeptin decreased the fluorescence signal detected compared to the untreated control (see table), which displayed a baseline level of cathepsin activity)
Related Product Information for Catb-2744 detection kit
Description: Green Cathepsin B Kit enables the quantitation and monitoring of intracellular cathepsin activity over time in vitro. The Rhodamine 110 Cathepsin B substrate reagent is a non-cytotoxic and membrane permeant substrate that fluoresces green upon cleavage by active cathepsin enzymes.
Test Principle: Rhodamine 110 Cathepsin B substrate utilizes the photostable green fluorophore, rhodamine 110. Rhodamine 110 cathepsin B substrate is comprised of rhodamine 110 coupled to two copies of the amino acid sequence, arginine-arginine (RR), which is the preferential target sequence for cathepsin B. When bi-substituted via amide linkage to two cathepsin B target peptide sequences, rhodamine110 is nonfluorescent. Following enzymatic cleavage at one or both arginine (R) amide linkage sites, the mono and non-substituted rhodamine 110 fluorophores generate green fluorescence when excited at 500 nm.
To use the Green Cathepsin B Assay, simply add the Rhodamine 110 Cathepsin B substrate [R110-(RR)2] directly to the cell culture media (or 1X Cellular Assay Buffer), incubate, and analyze. Because R110-(RR)2 is cell-permeant, it easily penetrates the cell membrane and the membranes of the internal cellular organelles - no lysis or permeabilization steps are required. R110-(RR)2 will enter the cell in a non-fluorescent state. If cathepsin enzymes are active, they will cleave off the two arginine-arginine cathepsin B targeting sequences and allow the rhodamine 110 fluorophore to become fluorescent upon excitation. By varying the duration and concentration of exposure to the R110-(RR)2 substrate, a picture can be obtained of the relative abundance and intracellular location of cathepsin enzymatic activity. Positive cells will fluoresce green, while negative cells will exhibit very low levels of background green fluorescence. There is no interference from pro-cathepsins forms of the enzymes. If the treatment or experimental condition stimulates cathepsin activity, cells containing elevated levels of cathepsin activity will appear brighter green than cells with lower levels of cathepsin activity.
Test Principle: Rhodamine 110 Cathepsin B substrate utilizes the photostable green fluorophore, rhodamine 110. Rhodamine 110 cathepsin B substrate is comprised of rhodamine 110 coupled to two copies of the amino acid sequence, arginine-arginine (RR), which is the preferential target sequence for cathepsin B. When bi-substituted via amide linkage to two cathepsin B target peptide sequences, rhodamine110 is nonfluorescent. Following enzymatic cleavage at one or both arginine (R) amide linkage sites, the mono and non-substituted rhodamine 110 fluorophores generate green fluorescence when excited at 500 nm.
To use the Green Cathepsin B Assay, simply add the Rhodamine 110 Cathepsin B substrate [R110-(RR)2] directly to the cell culture media (or 1X Cellular Assay Buffer), incubate, and analyze. Because R110-(RR)2 is cell-permeant, it easily penetrates the cell membrane and the membranes of the internal cellular organelles - no lysis or permeabilization steps are required. R110-(RR)2 will enter the cell in a non-fluorescent state. If cathepsin enzymes are active, they will cleave off the two arginine-arginine cathepsin B targeting sequences and allow the rhodamine 110 fluorophore to become fluorescent upon excitation. By varying the duration and concentration of exposure to the R110-(RR)2 substrate, a picture can be obtained of the relative abundance and intracellular location of cathepsin enzymatic activity. Positive cells will fluoresce green, while negative cells will exhibit very low levels of background green fluorescence. There is no interference from pro-cathepsins forms of the enzymes. If the treatment or experimental condition stimulates cathepsin activity, cells containing elevated levels of cathepsin activity will appear brighter green than cells with lower levels of cathepsin activity.
Product Categories/Family for Catb-2744 detection kit
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
NCBI Official Full Name
cathepsin B
NCBI Official Symbol
Catb-2744
NCBI Protein Information
cathepsin B
Protein Family
Similar Products
Product Notes
The Catb-2744 (Catalog #AAA225836) is a Detection Kit and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's Cathepsin B can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS), Immunofluorescence (IF). Researchers should empirically determine the suitability of the Catb-2744 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "Cathepsin B, Detection Kit" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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