Puromycin aminonucleoside (NSC 3056) is the aminonucleoside portion of the antibiotic puromycin, and used in nephrosis animal models. Puromycin aminonucleoside induces apoptosis. Puromycin aminonucleoside is a reversible inhibitor of dipeptidyl peptidase II and cytosol alanyl aminopeptidase. Puromycin aminonucleoside induces secretion of cell migrasome.
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Puromycin aminonucleoside Chemical Structure
CAS No. : 58-60-6
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Based on 22 publication(s) in Google Scholar
Puromycin aminonucleoside purchased from MedChemExpress. Usage Cited in:
Am J Transl Res. 2020 Jul 15;12(7):3512-3521.
[Abstract]
Expression of downstream proteins of the PCP pathway, such as ROCK1 and Rac1, is decreased in the Puromycin aminonucleoside (PAN) group and significantly enhanced by AS-IV treatment.
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Description
Puromycin aminonucleoside (NSC 3056) is the aminonucleoside portion of the antibiotic puromycin, and used in nephrosis animal models[1]. Puromycin aminonucleoside induces apoptosis[2]. Puromycin aminonucleoside is a reversible inhibitor of dipeptidyl peptidase II and cytosol alanyl aminopeptidase[3]. Puromycin aminonucleoside induces secretion of cell migrasome[4].
IC50 & Target
DPP-2
In Vitro
Puromycin aminonucleoside (NSC 3056) (30 μg/mL) markedly increases p53 protein levels in podocytes. Puromycin aminonucleoside (NSC 3056)-induced podocyte apoptosis is p53 dependent. Puromycin aminonucleoside (NSC 3056) induces podocyte apoptosis in a time-dependent manner[2]. The IC50 values for PMAT-expressing and vector-transfected cells are 48.9 and 122.1 μM, respectively, suggesting expression of PMAT-enhanced cell sensitivity to Puromycin aminonucleoside. Puromycin aminonucleoside (NSC 3056) (250 μM) is toxic to both PMAT-expressing and vector-transfected cells. Puromycin aminonucleoside (NSC 3056) uptake in PMAT-expressing cells is fourfold higher at pH 6.6 than that at pH 7.4[5].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Puromycin aminonucleoside Related Antibodies
In Vivo
Puromycin aminonucleoside (NSC 3056) (30 μg/mL) markedly increases p53 protein levels in podocytes. Puromycin aminonucleoside (NSC 3056)-induced podocyte apoptosis is p53 dependent. Puromycin aminonucleoside (NSC 3056) induces podocyte apoptosis in a time-dependent manner[2]. The IC50 values for PMAT-expressing and vector-transfected cells are 48.9 and 122.1 μM, respectively, suggesting expression of PMAT-enhanced cell sensitivity to Puromycin aminonucleoside. Puromycin aminonucleoside (NSC 3056) (250 μM) is toxic to both PMAT-expressing and vector-transfected cells. Puromycin aminonucleoside (NSC 3056) uptake in PMAT-expressing cells is fourfold higher at pH 6.6 than that at pH 7.4[5].
Induction of Nephrotic syndrome (NS) model[8]
Background
Puromycin aminonucleoside (PAN) induces cellular oxidant injury and increases in proteinuria, albumin (ALB), and creatinine clearance (Ccr) levels in the kidneys.
Specific Modeling Methods
Mice: Sprague Dawley rats •male •? 180-190 g Administration: 50 mg/kg • iv • single dose
Note
Modeling Indicators
Molecular changes: Induced increases in proteinuria, albumin (ALB), and Ccr levels.
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Room temperature in continental US; may vary elsewhere.
Stockage
Powder
-20°C
3 years
4°C
2 years
In solvent
-80°C
6 months
-20°C
1 month
Solvant et solubilité
In Vitro:
H2O : 40 mg/mL (135.91 mM; ultrasonic and warming and heat to 50°C)
DMSO : 25 mg/mL (84.94 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Preparing Stock Solutions
ConcentrationSolventMass
1 mg
5 mg
10 mg
1 mM
3.3978 mL
16.9889 mL
33.9778 mL
5 mM
0.6796 mL
3.3978 mL
6.7956 mL
10 mM
0.3398 mL
1.6989 mL
3.3978 mL
View the Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
*
Note: If you choose water as the stock solution, please dilute it to the working solution,
then filter and sterilize it with a 0.22 μm filter before use.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Protocol 1
Add each solvent one by one: 5% DMSO 40% PEG300 5% Tween-80 50% Saline
Solubility: ≥ 2.5 mg/mL (8.49 mM); Clear solution
Protocol 2
Add each solvent one by one: 5% DMSO 95% (20% SBE-β-CD in Saline)
Solubility: ≥ 2.5 mg/mL (8.49 mM); Clear solution
Protocol 3
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 4
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 5
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 6
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 900 μLCorn oil, and mix evenly.
Protocol 9
Add each solvent one by one: 10% DMSO 90% Corn Oil
This protocol yields a clear solution of ≥ 2.08 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (20.8 mg/mL) to 900 μLCorn oil, and mix evenly.
For the following dissolution methods, please prepare the working solution directly.
It is recommended to prepare fresh solutions and use them promptly within a short period of time. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution.
If precipitation or phase separation occurs during preparation,
heat and/or sonication can be used to aid dissolution.
Protocol 1
Add each solvent one by one: PBS
Solubility: 12.5 mg/mL (42.47 mM); Clear solution; Need ultrasonic
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
Dosage
mg/kg
Animal weight (per animal)
g
Dosing volume (per animal)
μL
Number of animals
Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Calculation results:
Working solution concentration:
mg/mL
This product has good water solubility, please refer to the measured solubility data in water/PBS/Saline for details.
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only.If necessary, please contact MedChemExpress (MCE).
[1]. Lacalle RA, et al. Cloning of the complete biosynthetic gene cluster for an aminonucleoside antibiotic, puromycin, and its regulated expression in heterologous hosts. EMBO J. 1992 Feb;11(2):785-92.
[Content Brief]
[2]. Wada T, et al. Prevents podocyte apoptosis induced by puromycin aminonucleoside: role of p53 and Bcl-2-related family proteins. J Am Soc Nephrol. 2005 Sep;16(9):2615-25.
[Content Brief]
[3]. Gong W, et al. Estrogen-related receptor-α mediates puromycin aminonucleoside-induced mesangial cell apoptosis and inflammatory injury. Am J Physiol Renal Physiol. 2019 May 1;316(5):F906-F913.
[Content Brief]
[4]. Ying Liu, et al. Podocyte-Released Migrasomes in Urine Serve as an Indicator for Early Podocyte Injury. Kidney Dis (Basel). 2020 Nov;6(6):422-433.
[Content Brief]
[5]. Xia L, et al. Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleosidenephrotoxicity. Am J Physiol Renal Physiol. 2009 Jun;296(6):F1307-13.
[Content Brief]
[6]. Kawakami H, et al. Dynamics of absolute amount of nephrin in a single podocyte in puromycin aminonucleoside nephrosis rats calculated by quantitative glomerular proteomics approach with selected reaction monitoring mode. Nephrol Dial Transplant. 2012 Apr;
[Content Brief]
[7]. Nosaka K, et al. An adenosine deaminase inhibitor prevents puromycin aminonucleoside nephrotoxicity. Free Radic Biol Med 1997 ;22 (4): 597-605.
[Content Brief]
Test cellulaire
[4]
Cells are seeded in MEM with 10% FBS on 96-well plates at a density of 5,000 cells/well. After appr 48-h incubation (appr 40-50% confluence), cells are changed to fresh growth medium containing Puromycin aminonucleoside (NSC 3056) at various concentrations. For the protection experiment, cells are incubated in medium containing 250 μM Puromycin aminonucleoside (NSC 3056) with or without the PMAT inhibitor decynium-22 (2 μM). After a total of 72-h incubation in a 95% O2 incubator at 37°C, cells are washed and the plates. The IC50 values are determined by fitting the cell growth data to the following model using nonlinear regression (WinNonLin version 3.2): S=Smax − [Smax − S0] × [Cγ/(Cγ + IC50γ)], where S is the cell survival expressed as percentage of the optical density to untreated control cells, Smax is the maximal cell survival, S0 is the lowest residual cell survival at the high drug concentration, C is Puromycin aminonucleoside concentration, γ is the Hill coefficient, and IC50 is the Puromycin aminonucleoside concentration leading to half-maximal cell survival. Five to six determinations are carried out within each experiment, and four independent experiments are performed.
MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Administration animale
[5]
Male F344 rats at 11 weeks of age are purchased from JaPuromycin aminonucleoside SLC. Normal rats and a Puromycin aminonucleoside nephrosis model are used in the present study. Puromycin aminonucleoside (NSC 3056) nephrosis is induced in rats by a single intravenous injection of Puromycin aminonucleoside at a dose of 8 mg/100 g body weight in saline. Control animals receive an identical volume of saline. Nephrotic rats (n=6 per group) are studied at Days 4 and 7 after the Puromycin aminonucleoside injection.
MCE n'a pas confirmé de manière indépendante l'exactitude de ces méthodes. Ils sont pour référence seulement.
Références
[1]. Lacalle RA, et al. Cloning of the complete biosynthetic gene cluster for an aminonucleoside antibiotic, puromycin, and its regulated expression in heterologous hosts. EMBO J. 1992 Feb;11(2):785-92.
[Content Brief]
[2]. Wada T, et al. Prevents podocyte apoptosis induced by puromycin aminonucleoside: role of p53 and Bcl-2-related family proteins. J Am Soc Nephrol. 2005 Sep;16(9):2615-25.
[Content Brief]
[3]. Gong W, et al. Estrogen-related receptor-α mediates puromycin aminonucleoside-induced mesangial cell apoptosis and inflammatory injury. Am J Physiol Renal Physiol. 2019 May 1;316(5):F906-F913.
[Content Brief]
[4]. Ying Liu, et al. Podocyte-Released Migrasomes in Urine Serve as an Indicator for Early Podocyte Injury. Kidney Dis (Basel). 2020 Nov;6(6):422-433.
[Content Brief]
[5]. Xia L, et al. Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleosidenephrotoxicity. Am J Physiol Renal Physiol. 2009 Jun;296(6):F1307-13.
[Content Brief]
[6]. Kawakami H, et al. Dynamics of absolute amount of nephrin in a single podocyte in puromycin aminonucleoside nephrosis rats calculated by quantitative glomerular proteomics approach with selected reaction monitoring mode. Nephrol Dial Transplant. 2012 Apr;
[Content Brief]
[7]. Nosaka K, et al. An adenosine deaminase inhibitor prevents puromycin aminonucleoside nephrotoxicity. Free Radic Biol Med 1997 ;22 (4): 597-605.
[Content Brief]
[1]. Lacalle RA, et al. Cloning of the complete biosynthetic gene cluster for an aminonucleoside antibiotic, puromycin, and its regulated expression in heterologous hosts. EMBO J. 1992 Feb;11(2):785-92.
[2]. Wada T, et al. Prevents podocyte apoptosis induced by puromycin aminonucleoside: role of p53 and Bcl-2-related family proteins. J Am Soc Nephrol. 2005 Sep;16(9):2615-25.
[3]. Gong W, et al. Estrogen-related receptor-α mediates puromycin aminonucleoside-induced mesangial cell apoptosis and inflammatory injury. Am J Physiol Renal Physiol. 2019 May 1;316(5):F906-F913.
[4]. Ying Liu, et al. Podocyte-Released Migrasomes in Urine Serve as an Indicator for Early Podocyte Injury. Kidney Dis (Basel). 2020 Nov;6(6):422-433.
[5]. Xia L, et al. Podocyte-specific expression of organic cation transporter PMAT: implication in puromycin aminonucleosidenephrotoxicity. Am J Physiol Renal Physiol. 2009 Jun;296(6):F1307-13.
[6]. Kawakami H, et al. Dynamics of absolute amount of nephrin in a single podocyte in puromycin aminonucleoside nephrosis rats calculated by quantitative glomerular proteomics approach with selected reaction monitoring mode. Nephrol Dial Transplant. 2012 Apr;
[7]. Nosaka K, et al. An adenosine deaminase inhibitor prevents puromycin aminonucleoside nephrotoxicity. Free Radic Biol Med 1997 ;22 (4): 597-605.
Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Optional Solvent
ConcentrationSolventMass
1 mg
5 mg
10 mg
25 mg
DMSO / H2O
1 mM
3.3978 mL
16.9889 mL
33.9778 mL
84.9444 mL
5 mM
0.6796 mL
3.3978 mL
6.7956 mL
16.9889 mL
10 mM
0.3398 mL
1.6989 mL
3.3978 mL
8.4944 mL
15 mM
0.2265 mL
1.1326 mL
2.2652 mL
5.6630 mL
20 mM
0.1699 mL
0.8494 mL
1.6989 mL
4.2472 mL
25 mM
0.1359 mL
0.6796 mL
1.3591 mL
3.3978 mL
30 mM
0.1133 mL
0.5663 mL
1.1326 mL
2.8315 mL
40 mM
0.0849 mL
0.4247 mL
0.8494 mL
2.1236 mL
50 mM
0.0680 mL
0.3398 mL
0.6796 mL
1.6989 mL
60 mM
0.0566 mL
0.2831 mL
0.5663 mL
1.4157 mL
80 mM
0.0425 mL
0.2124 mL
0.4247 mL
1.0618 mL
H2O
100 mM
0.0340 mL
0.1699 mL
0.3398 mL
0.8494 mL
*
Note: If you choose water as the stock solution, please dilute it to the working solution,
then filter and sterilize it with a 0.22 μm filter before use.
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.
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