| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
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| Targets |
Puromycin-bis(PEG2-amide)-Biotin shares the same target as puromycin, which is the ribosomal A site. It binds to the ribosome and mimics the 3' end of aminoacyl-tRNA, leading to premature termination of protein synthesis.
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|---|---|
| ln Vitro |
Puromycin-bis(PEG2-amide)-Biotin efficiently incorporated into nascent polypeptides in cultured cells, as detected by streptavidin-based pull-down assays and mass spectrometry. This demonstrated its ability to label newly synthesized proteins for subsequent purification and analysis [1]
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| ln Vivo |
In C57BL/6 mice, intraperitoneal injection of Puromycin-bis(PEG2-amide)-Biotin (50 mg/kg) resulted in labeling of newly synthesized proteins in multiple tissues. Tissues with high protein synthesis rates, such as the intestinal epithelium and bone marrow, showed strong biotin signals detected by streptavidin-fluorophore conjugation. Moderate signals were observed in the liver and kidney, reflecting baseline protein synthesis levels [1]
The biotinylated protein signal peaked at 1 hour post-injection and remained detectable for up to 4 hours, enabling temporal analysis of in vivo protein synthesis dynamics [1] |
| Enzyme Assay |
Ribosome Binding Assay: Radiolabeled Puromycin-bis(PEG2-amide)-Biotin was incubated with purified ribosomes in buffer containing Mg²⁺ and ATP. Binding was measured by nitrocellulose filter retention, showing specific interaction with ribosomes. Competition assays with unlabeled puromycin confirmed the binding specificity [1]
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| Cell Assay |
rotein Labeling and Purification: Cells were treated with Puromycin-bis(PEG2-amide)-Biotin (1–10 μM) for 30–60 minutes, followed by lysis and streptavidin bead purification. Western blot analysis confirmed the enrichment of biotinylated proteins, indicating successful labeling of newly synthesized polypeptides [1]
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| Animal Protocol |
- In Vivo Protein Labeling in Mice: Puromycin-bis(PEG2-amide)-Biotin (50 mg/kg) was administered intraperitoneally to C57BL/6 mice. After 1 hour, tissues were harvested and processed for streptavidin-based detection. Fluorescence imaging showed labeling in highly proliferative tissues like the intestine and bone marrow [1]
- Drug Formulation: Puromycin-bis(PEG2-amide)-Biotin was dissolved in sterile DMSO and diluted with PBS to a final concentration of 10 mg/mL before injection [1]
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| References |
[1]. Yungui Yang, et al. Single-cell translatome sequencing method. WO2023284076A1.
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| Additional Infomation |
- Puromycin-bis(PEG2-amide)-Biotin is a modified puromycin analog with two PEG2-amide linkers and a biotin moiety, enabling bioorthogonal labeling of newly synthesized proteins for downstream applications such as proteomics and single-cell translatome sequencing [1]
- The compound combines the protein synthesis inhibitory activity of puromycin with the high-affinity binding of biotin to streptavidin, allowing efficient capture and analysis of nascent polypeptides in complex biological samples [1]
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| Molecular Formula |
C46H71N13O13S
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|---|---|
| Molecular Weight |
1046.20
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| CAS # |
2379889-82-2
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| Appearance |
White to off-white solid powder
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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 | 0.9558 mL | 4.7792 mL | 9.5584 mL | |
| 5 mM | 0.1912 mL | 0.9558 mL | 1.9117 mL | |
| 10 mM | 0.0956 mL | 0.4779 mL | 0.9558 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.
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