| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
Fmoc-Ala-Glu-Gln-Lys-NH2 is an inactive control peptide and does not bind to a specific biological target. The active control, Fmoc-Ala-Glu-Asn-Lys-NH2 (AENK), targets and blocks the proteolysis of UNC5C (Uncoordinated-5C, a netrin receptor involved in axon guidance and tumor suppression). The substitution of Asn with Gln in AEQK eliminates this inhibitory activity.
|
|---|---|
| ln Vitro |
The proteolysis of UNC5C is unaffected by Fmoc-Ala-Glu-Gln-Lys-NH2(45 min; pH=6), whereas Fmoc-Ala-Glu-Asn-Lys-NH2(AENK) exhibits inhibition[1].
Fmoc-Ala-Glu-Gln-Lys-NH2 has no intrinsic biological activity and serves as a negative control for the active peptide inhibitor AENK. In vitro, the active peptide AENK blocks the proteolysis of UNC5C protein. AEQK, with Gln replacing Asn, does not affect UNC5C proteolysis and is used to confirm that effects observed with AENK are sequence-specific. The control peptide is used to rule out off-target or non-specific effects. |
| ln Vivo |
In vivo, Fmoc-Ala-Glu-Gln-Lys-NH2 has no biological activity and is not intended for in vivo use as a therapeutic. It is used as a control in research studies to validate that the effects of the active peptide AENK on UNC5C proteolysis and downstream pathways are specific. The active peptide AENK has been studied in the context of UNC5C biology, which is involved in neuronal development and cancer suppression. AEQK serves as a specificity control in such studies.
|
| Enzyme Assay |
Cell-free binding and enzyme assays are not typically performed with the inactive control peptide. For validation of the active peptide's inhibitory mechanism, a cell-free proteolysis assay can be set up: incubate purified UNC5C protein or a UNC5C-derived peptide substrate (1-10 uM) with the relevant protease (e.g., calpain or other cysteine protease) in reaction buffer (50 mM Tris-HCl, pH 7.4, 2 mM CaCl2, 5 mM DTT) with or without Fmoc-Ala-Glu-Gln-Lys-NH2 (1-100 uM) as a negative control. Use the active peptide AENK as a positive control. Incubate at 37degC for 1-4 hours. Analyze by SDS-PAGE and Coomassie blue staining or by LC-MS to monitor substrate degradation. The inactive control should not inhibit proteolysis.
|
| Cell Assay |
Culture UNC5C-expressing cell lines (e.g., HEK293T transfected with UNC5C, or certain neuronal or cancer cell lines) in DMEM with 10% FBS. Treat cells with Fmoc-Ala-Glu-Gln-Lys-NH2 (inactive control) or Fmoc-Ala-Glu-Asn-Lys-NH2 (active AENK) at concentrations of 1-100 uM for 6-24 hours. Assess UNC5C protein levels by Western blot using anti-UNC5C antibody. Measure downstream signaling pathways (e.g., p53, pro-apoptotic markers, or axon guidance markers) as appropriate. The inactive control should not alter UNC5C levels. Perform cell viability assays (MTT, LDH) to confirm lack of non-specific cytotoxicity. Co-treat with proteasome inhibitor MG132 (10 uM) to block proteasomal degradation and confirm that AENK's effect is on proteolysis, not transcription. Use a scrambled peptide control in parallel for additional specificity confirmation. For cellular uptake studies, use fluorescently labeled versions (e.g., FITC-AEQK) and visualize by confocal microscopy. Quantify intracellular fluorescence by flow cytometry. Compare uptake efficiency between the active and inactive peptides to rule out differences in cellular penetration.
|
| Animal Protocol |
Animal studies are not typically performed with the inactive control peptide. For studies of the active AENK peptide in vivo (e.g., in tumor xenograft models or neurological disease models), administer AENK by intravenous or intraperitoneal injection at doses of 1-10 mg/kg. Include a control group receiving Fmoc-Ala-Glu-Gln-Lys-NH2 at the same dose as a negative control. After treatment, harvest target tissues (e.g., tumors, brain, spinal cord). Process tissues for Western blot analysis of UNC5C levels, immunohistochemistry, and assessment of downstream signaling. The inactive control group should show no significant difference from vehicle-treated controls, confirming that any effects observed with AENK are due to specific UNC5C proteolysis blockade. Monitor body weight, organ weights, and general health as safety readouts. Collect plasma for PK analysis and to measure cytokine levels if inflammation is a concern. Use inactive control to assess non-specific peptide effects on immune activation. For survival studies in disease models, treat with AENK, inactive control, or vehicle, and plot Kaplan-Meier survival curves. The inactive control should not affect survival compared to vehicle.
|
| ADME/Pharmacokinetics |
Fmoc-Ala-Glu-Gln-Lys-NH2 is a tetrapeptide (MW 695.76, formula C34H45N7O9). The Fmoc (9-fluorenylmethoxycarbonyl) group protects the N-terminus and is acid-labile. The C-terminus is amidated (NH2). As a protected peptide, it is not intended for in vivo administration as a therapeutic; it is a research tool for specificity control. The compound is soluble in DMSO. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month). Protect from light and moisture. The amide (-NH2) C-terminus increases stability toward carboxypeptidase degradation. The substitution of Asn (in active peptide) with Gln (in inactive control) changes the hydrogen bonding capacity and side-chain length, eliminating biological activity. The purity is typically ≥98% by HPLC. For in vitro use, prepare stock solutions in DMSO (10-100 mM) and dilute in cell culture medium (final DMSO <0.1%). For in vivo control studies, dissolve in saline or PBS with pH adjustment if necessary. Filter sterilize before injection.
|
| Toxicity/Toxicokinetics |
The inactive control peptide is considered safe for laboratory use under standard chemical safety guidelines. It is not intended for human consumption. As a peptide, it is expected to have low acute toxicity. The Fmoc protecting group is not cleaved in vivo, and the intact peptide is likely to be metabolized by proteases. No genotoxicity or carcinogenicity data are available. Standard laboratory precautions (gloves, lab coat, safety glasses) should be used. Avoid inhalation, ingestion, and skin contact. For large-scale synthesis, follow guidelines for handling protected peptides. Dispose of waste according to institutional chemical waste protocols. In case of accidental skin contact, wash with soap and water; if eye contact, rinse with water for 15 minutes. If ingested, seek medical attention. This compound is not classified as hazardous for transport under UN regulations.
|
| References | |
| Additional Infomation |
Fmoc-Ala-Glu-Gln-Lys-NH2 (AEQK) is a tetrapeptide that serves as an inactive control for the peptide inhibitor Fmoc-Ala-Glu-Asn-Lys-NH2 (AENK). AENK blocks proteolysis of UNC5C protein, which is involved in axon guidance (netrin-1 receptor) and functions as a dependence receptor inducing apoptosis in the absence of its ligand. UNC5C is also a tumor suppressor in certain cancers. The AEQK control peptide contains glutamine (Gln) instead of asparagine (Asn) at the third position, which eliminates inhibitory activity. This control is essential for validating that the effects of AENK are due to specific inhibition of UNC5C proteolysis. The compound is exclusively for research use and not for diagnostic or therapeutic applications.
|
| Molecular Formula |
C34H45N7O9
|
|---|---|
| Molecular Weight |
695.76
|
| Appearance |
White to off-white 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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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) |
DMSO :~3.57 mg/mL (~5.13 mM)
H2O :< 0.1 mg/mL |
|---|---|
| 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 | 1.4373 mL | 7.1864 mL | 14.3728 mL | |
| 5 mM | 0.2875 mL | 1.4373 mL | 2.8746 mL | |
| 10 mM | 0.1437 mL | 0.7186 mL | 1.4373 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
