| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Mutant TIR1 F-box proteins (e.g., AtTIR1(F79G)). The AID2 system is a two-component system where the target protein is fused to a small degron tag. In the presence of 5-Ph-IAA, the plant-derived F-box protein TIR1 binds to both the degron tag and the ligand, polyubiquitinating the target protein and leading to its degradation by the proteasome. 5-Ph-IAA-AM is the cell-permeable ester prodrug of the active ligand.
|
|---|---|
| ln Vitro |
5-Ph-IAA-AM is the membrane-permeable ester analog of the active degrader, 5-Ph-IAA. It has no direct activity until it is inside the cell. Once taken up, cellular esterases hydrolyze the AM ester to release the active 5-Ph-IAA. Active 5-Ph-IAA then binds to the engineered TIR1 F-box protein with high affinity (DC50 at 6h = 17.0 nM), promoting selective ubiquitination and degradation of target degron-tagged proteins.
|
| ln Vivo |
In laid C. elegans embryos, the use of 5-Ph-IAA-AM affords enhanced protein degradation of tagged proteins compared to the parent compound 5-Ph-IAA. This is because the AM-ester form enables efficient penetration of the eggshell, allowing temporally controlled protein knockdown during embryogenesis and development. It has been used to dissect the function of essential proteins in early development.
|
| Enzyme Assay |
A standard assay for the AID2 system involves expressing a degron-tagged protein of interest (e.g., a GFP-fusion) and the AtTIR1(F79G) protein in cells. Protein degradation is triggered by adding 5-Ph-IAA-AM to the culture medium. The efficiency and kinetics of degradation are monitored over time by western blot or by fluorescence microscopy for fluorescently tagged proteins, using the parent compound 5-Ph-IAA as a control.
|
| Cell Assay |
In a typical cellular assay, a mammalian cell line (e.g., HeLa, HEK293T) is engineered to stably express the AtTIR1(F79G) protein and the degron-tagged target protein. Cells are seeded in multi-well plates and treated with varying concentrations of 5-Ph-IAA-AM (or the active 5-Ph-IAA) for a defined time (e.g., 1-24 hours). Cells are then lysed, and the remaining target protein is quantified by western blot to calculate the half-maximal degradation concentration (DC50). Cell viability is also monitored.
|
| Animal Protocol |
The AID2 system can be used in vivo in model organisms like C. elegans. Transgenic C. elegans lines are generated that express the AtTIR1(F79G) protein and the degron-tagged target protein. For experiments with embryos, 5-Ph-IAA-AM is added directly to the culture medium or growth plates because it can penetrate the eggshell. The effect of protein depletion on embryogenesis and development is then observed over time using microscopy.
|
| ADME/Pharmacokinetics |
5-Ph-IAA-AM is a small, cell-permeable prodrug. Its pharmacokinetic properties are not fully characterized, but it is designed to be rapidly taken up by cells and cleaved by intracellular esterases to release the active 5-Ph-IAA. The active acid, 5-Ph-IAA, has a molecular weight of 267.28 and a half-life (T1/2) of 62.3 minutes for the degradation reaction. The AM ester is less stable and hydrolyzes quickly in aqueous solution.
|
| Toxicity/Toxicokinetics |
Specific toxicity data for 5-Ph-IAA-AM is not available, but it is considered a tool for chemical biology and is generally used at low concentrations to induce rapid protein degradation. In the AID2 system, the ligand itself is not typically toxic, as it only drives the degradation of the tagged target. However, toxicity could arise from the loss of an essential protein. For research use, it is handled as a potentially hazardous chemical using standard safety precautions.
|
| References |
|
| Additional Infomation |
5-Ph-IAA-AM is an organic molecular entity.
5-Ph-IAA-AM is a chemical biology tool used for the auxin-inducible degron 2 (AID2) system, an improved version of the original AID system. Compared to the original auxin (IAA) which can induce off-target effects through endogenous auxin signaling pathways, the 5-Ph-IAA/AtTIR1(F79G) pair is orthogonal with no known off-target effects in animal or yeast cells. This system enables rapid, reversible, and highly specific conditional protein depletion, providing a powerful tool for studying protein function in living cells and whole organisms. |
| Molecular Formula |
C19H17NO4
|
|---|---|
| Molecular Weight |
323.34
|
| Exact Mass |
323.116
|
| CAS # |
2990123-10-7
|
| PubChem CID |
162641049
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
4
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
24
|
| Complexity |
447
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CC(=O)OCOC(=O)CC1=CNC2=C1C=C(C=C2)C3=CC=CC=C3
|
| InChi Key |
FGVMSTLYCNILJO-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C19H17NO4/c1-13(21)23-12-24-19(22)10-16-11-20-18-8-7-15(9-17(16)18)14-5-3-2-4-6-14/h2-9,11,20H,10,12H2,1H3
|
| Chemical Name |
acetyloxymethyl 2-(5-phenyl-1H-indol-3-yl)acetate
|
| 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 |
| 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: 100 mg/mL (309.27 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.73 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.73 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.73 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0927 mL | 15.4636 mL | 30.9272 mL | |
| 5 mM | 0.6185 mL | 3.0927 mL | 6.1854 mL | |
| 10 mM | 0.3093 mL | 1.5464 mL | 3.0927 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
