| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
PEGs
Iodoacetamide-PEG5-azide does not target a biological receptor; it is a chemical crosslinker. The iodoacetamide group reacts specifically with free thiols (cysteine residues) to form stable thioether bonds. The azide group participates in copper-catalyzed or copper-free click chemistry with alkynes. The PEG5 spacer enhances water solubility and reduces steric hindrance. |
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| ln Vitro |
A linker separates the two ligands that make up PROTACs; one ligand is for an E3 ubiquitin ligase, and the other is for the target protein. Target proteins are selectively degraded by PROTACs by taking advantage of the intracellular ubiquitin-proteasome system[1].
In cell-free assays, the reactivity of iodoacetamide-PEG5-azide is tested with thiol-containing compounds (e.g., glutathione, cysteine) and alkyne-bearing molecules. The reaction with thiols proceeds at pH 7-8, forming a thioether. The azide group reacts with alkynes in the presence of copper catalyst or with strained cyclooctynes. Reaction progress is monitored by LC-MS or HPLC. No intrinsic biological activity is present. |
| ln Vivo |
Iodoacetamide-PEG5-azide is used to label surface or intracellular thiols in live or fixed cells. Cells (e.g., 10⁶ cells/mL) are treated with 1-100 uM of the compound in PBS (pH 7.4) for 30-60 minutes at 4degC or room temperature. After washing, the azide group can be further reacted with a fluorescent alkyne via click chemistry (e.g., Alexa Fluor 488-alkyne) to visualize thiol-containing proteins. The compound is not toxic at these concentrations.
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| Enzyme Assay |
For thiol reactivity, a cell-free assay: a model thiol (e.g., reduced glutathione, 100 uM) is incubated with Iodoacetamide-PEG5-azide (100 uM) in PBS (pH 7.4) at 37degC. Aliquots are taken at 0, 15, 30, 60, 120 minutes and analyzed by LC-MS to quantify the adduct. The reaction is typically complete within 1 hour. For azide activity, the purified thiol-conjugated product is then reacted with a cyclooctyne (e.g., DBCO-Cy5) and analyzed by SDS-PAGE fluorescence.
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| Cell Assay |
Cells are treated with Iodoacetamide-PEG5-azide (10-100 uM) for 30-60 minutes to alkylate free thiols. After washing, cells are incubated with an alkyne-fluorophore (e.g., alkyne-AF488, 10 uM) plus CuSO4/THPTA/sodium ascorbate for 30 minutes at room temperature (for CuAAC) or with a cyclooctyne-fluorophore for copper-free labeling. Cells are then washed and analyzed by flow cytometry or fluorescence microscopy. Controls include pre-blocking thiols with N-ethylmaleimide.
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| Animal Protocol |
Iodoacetamide-PEG5-azide can be used in vivo for pretargeted imaging or drug delivery. A thiol-reactive probe is first injected (e.g., intravenously) to alkylate free thiols on tumor surface proteins. After a washout period, a radiolabeled or fluorescent alkyne probe is injected to react with the azide via click chemistry in vivo. This two-step pretargeting approach reduces background. Doses of 1-10 mg/kg have been used in mice. Biodistribution is assessed by imaging or tissue counting.
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| ADME/Pharmacokinetics |
The compound has a molecular weight of approximately 450-550 Da. It is soluble in water and organic solvents. The iodoacetamide group is sensitive to light and hydrolysis; store at -20degC under inert atmosphere, protected from light. The PEG5 linker provides hydrophilicity and reduces non-specific binding. The azide group is stable under biological conditions. The plasma half-life of the free linker is short (minutes); but when conjugated to a protein, PK is governed by the protein.
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| Toxicity/Toxicokinetics |
Iodoacetamide-PEG5-azide is an alkylating agent and should be handled with care. It may cause skin and eye irritation. At typical working concentrations (≤100 uM), no significant cytotoxicity has been observed in short-term assays. In animal studies at ≤10 mg/kg, no overt toxicity was reported. The compound is for research use only; no clinical safety data are available. Use appropriate PPE.
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| References | |
| Additional Infomation |
Iodoacetamide-PEG5-azide is a multifunctional linker for bioconjugation and chemical biology. It enables site-specific labeling of thiol-containing proteins followed by click chemistry attachment of reporters or payloads. The PEG5 spacer reduces steric hindrance and improves solubility. This compound is widely used in proteomics (e.g., enrichment of thiol-containing peptides), imaging, and drug delivery research. Not for human therapeutic use. Store desiccated at -20degC.
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| Molecular Formula |
C14H27IN4O6
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|---|---|
| Molecular Weight |
474.29
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| Appearance |
Light yellow to yellow liquid
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| 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 | 2.1084 mL | 10.5421 mL | 21.0841 mL | |
| 5 mM | 0.4217 mL | 2.1084 mL | 4.2168 mL | |
| 10 mM | 0.2108 mL | 1.0542 mL | 2.1084 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
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