| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Cathepsin D and E FRET Substrate acetate is a substrate for cathepsin D and cathepsin E, both aspartic proteases. It is not cleaved by cathepsins B, H, or I. Cleavage occurs at the Phe-Phe amide bond within the peptide sequence. This substrate is used as a tool to measure the enzymatic activity of cathepsin D and E in various biological samples.
|
|---|---|
| ln Vitro |
This FRET substrate has no intrinsic inhibitory or toxic activity on cells. It is used as a probe to measure cathepsin D and E activity in biological fluids, cell lysates, or tissue homogenates. Upon cleavage by active cathepsin D or E, the fluorophore (MOCAc) is separated from the quencher [Lys(Dnp)], resulting in a dose-dependent increase in fluorescence that is proportional to enzyme activity.
|
| ln Vivo |
Cathepsin D and E FRET Substrate acetate is not used for direct in vivo administration. It is primarily a biochemical tool for in vitro enzyme activity assays. However, it can be used in ex vivo assays on tissue homogenates or biological fluids collected from animal models to assess cathepsin activity changes related to disease (e.g., cancer, neurodegeneration).
|
| Enzyme Assay |
For in vitro enzyme assays, the FRET substrate is dissolved in assay buffer (e.g., 0.1 M sodium acetate, pH 4.0-5.5 for cathepsin D/E optimal activity) to a final concentration of 1-10 microM. The reaction is initiated by adding recombinant or purified cathepsin enzyme, and fluorescence (excitation 328 nm, emission 393 nm) is measured continuously for 30-60 minutes at 37degC. Inhibition studies are performed by pre-incubating the enzyme with inhibitors.
|
| Cell Assay |
For cellular assays, cells are lysed in a buffer containing 0.1% Triton X-100 in 0.1 M sodium acetate (pH 4.0) to activate lysosomal cathepsins. The lysate is clarified by centrifugation and incubated with the FRET substrate (1-10 microM) for 30-120 minutes at 37degC. Fluorescence is measured in a plate reader. Specific cathepsin activity is determined by using selective inhibitors (e.g., pepstatin A for cathepsin D/E).
|
| Animal Protocol |
For ex vivo analysis, tissues from animal models are homogenized in lysis buffer. The homogenate is centrifuged, and the supernatant is collected. Protein concentration is determined, and equal amounts of protein are incubated with the FRET substrate (1-10 microM) in activity buffer (pH 4.0) for 60 minutes at 37degC. Fluorescence is measured, and activity is normalized to protein concentration. Pepstatin A (1 microM) is used as a control for specificity.
|
| ADME/Pharmacokinetics |
As a biochemical substrate, the acetate salt form is highly water-soluble, which facilitates use in aqueous assay buffers. It has no independent pharmacokinetic properties as it is not administered in vivo. For in vitro assays, the substrate is stable in solution at 4degC for short-term use (hours) and should be stored as a powder at -20degC for long-term stability.
|
| Toxicity/Toxicokinetics |
The acetate salt form is considered safe for laboratory use under standard safety guidelines. The substrate is not intended for human or animal consumption and should be handled as a potentially hazardous chemical (avoid ingestion, inhalation, and skin contact). It is for research use only. No significant toxicity is expected at the low concentrations used in enzymatic assays (typically 1-10 microM).
|
| References | |
| Additional Infomation |
Cathepsin D and E FRET Substrate acetate is a fluorogenic substrate with the sequence MOCAc-Gly-Lys-Pro-Ile-Leu-Phe-Phe-Arg-Leu-Lys(Dnp)-D-Arg-NH2. Upon cleavage by cathepsin D or E, the MOCAc fluorophore (ex 328/em 393 nm) is separated from the Dnp quencher. It is a valuable tool for studying these proteases in cancer, neurodegeneration, and inflammation. Purity is ≥98% by HPLC.
|
| Molecular Formula |
C87H126N22O21
|
|---|---|
| Molecular Weight |
1816.15
|
| Related CAS # |
Cathepsin D and E FRET Substrate;839730-93-7
|
| Appearance |
Yellow to orange solid powder
|
| HS Tariff Code |
2934.99.9001
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O :~10 mg/mL (~5.51 mM)
|
|---|---|
| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 0.5506 mL | 2.7531 mL | 5.5062 mL | |
| 5 mM | 0.1101 mL | 0.5506 mL | 1.1012 mL | |
| 10 mM | 0.0551 mL | 0.2753 mL | 0.5506 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
