Pitstop 2d
Based on 1 Customer Validation
Pitstop 2d is a clathrin (Clathrin) terminal domain (TD) inhibitor with an IC50 of 1.7 μM. Pitstop 2d occupies the Clathrin box binding site within the clathrin TD, blocking the binding of endocytic protein ligands such as Epsin to clathrin. Pitstop 2d inhibits clathrin-mediated endocytosis (Endocytosis) and the cellular entry of vesicular stomatitis virus (VSV). Pitstop 2d has low cytotoxicity, does not disrupt the nuclear permeability barrier, and does not inhibit clathrin-independent endocytosis. Pitstop 2d can be used in studies related to endocytosis and vesicular stomatitis virus infection.
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- Purity: 97.11%
- 화학식: C26H18N2O4S2
- 분자량:486.56
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보관:
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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용액&용해도
- 순도&문서
- References
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Help & FAQs
Biological Activity
제품 설명
IC50 & Target
IC50: <2 μM (Clathrin TD)[1]
In Vitro
Pitstop 2d potently inhibits the interaction between purified clathrin terminal domain and amphiphysin B/C domain in cell-free ELISA assays, with an IC50 of 1.7 μM[1].
Pitstop 2d (25-100 μM; 8-20 h) exhibits only extremely low cytotoxicity in HEK293 and Cos7 cells[1].
Pitstop 2d (30 μM; 15 min) does not disrupt the nuclear permeability barrier in HeLa cells[1].
Pitstop 2d (13-45 μM; 15 min preincubation followed by 15 min transferrin uptake; 4 h washout) inhibits clathrin-mediated transferrin endocytosis in HeLa, HEK293T and Cos7 cells, with an IC50 of 13 μM in HeLa cells, and its inhibitory effect is reversible after washout[1].
Pitstop 2d significantly reduces the proportion of dynamic clathrin-coated pits in Cos7 cells expressing eGFP-clathrin light chain α, shifting CCP dynamics toward long-lived structures[1].
Pitstop 2d interferes with clathrin dynamics in the perinuclear region of Cos7 cells expressing eGFP-clathrin light chain α, reducing the maximum fluorescence recovery rate after photobleaching to 37.6%[1].
Pitstop 2d (45 μM; 15 min preincubation followed by 60 min dextran uptake) does not inhibit fluid-phase endocytosis of Alexa488-labeled dextran in HeLa cells[1].
Pitstop 2d binds to the clathrin box site of the purified clathrin terminal domain in an extended conformation, forming a specific and stable interaction[1].
Pitstop 2d (100 μM; 1 h) reduces the binding level between clathrin and Epsin 1 in Cos7 cells expressing eGFP-clathrin light chain α, while increasing the binding level between clathrin and SNX9, but has no effect on most other early-acting endocytic proteins[1].
Pitstop 2d (15 min) increases the total number of clathrin-coated structures on the plasma membrane of BS-C-1 cells, causes the accumulation of shallow clathrin-coated pits, but does not alter the diameter of free clathrin-coated vesicles[1].
Pitstop 2d (20 μM; 15 min preincubation followed by 1 h virus infection) inhibits over 60% of VSV entry into Vero-E6 cells, confirming that clathrin-mediated endocytosis (CME) is the primary pathway for efficient VSV cellular entry[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:HEK293, Cos7 cells
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Concentration:25, 50 and 100 μM
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Incubation Time:8 and 20 h
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Result:Exhibited low cytotoxicity, with minimal LDH release detected across all tested concentrations and incubation times, in contrast to the high cytotoxicity of Pitstop 2.
Quantified cytotoxicity as 1.53% relative to maximal LDH release induced by Triton X-100.
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Cell Line:Cos7 cells expressing eGFP-clathrin light chain α
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Concentration:100 μM
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Incubation Time:1 h
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Result:Reduced the binding level of clathrin to Epsin 1.
Increased the binding level of clathrin to SNX9.
Had no effect on most other early-acting endocytic proteins.
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Cell Line:HeLa cells
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Concentration:30 μM
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Incubation Time:15 min
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Result:Did not disrupt the nuclear permeability barrier
Chemical Information
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Appearance Solid
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분자량 486.56
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화학식 C26H18N2O4S2
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Color White to yellow
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SMILES
O=S(C1=CC2=C(C=C1)C=CC=C2)(NC(S/C3=C\C4=C(C=CC=C4)OC5=CC=CC=C5)=NC3=O)=O
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선적
Room temperature in continental US; may vary elsewhere.
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보관
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
용액&용해도
In Vitro:
DMSO : 100 mg/mL (205.52 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- 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.
Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (5.14 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (25.0 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.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
순도&문서
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Data Sheet (283 KB)
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SDS (252 KB)
- English - EN (252 KB)
- Français - FR (252 KB)
- Deutsch - DE (252 KB)
- Norwegian - NO (252 KB)
- Español - ES (252 KB)
- Swedish - SV (252 KB)
- Italian - IT (252 KB)
- Korean - KR (252 KB)
- Portuguese - PT (252 KB)
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Handling Instructions (2659 KB)
References
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 (sealed storage, away from moisture). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 2.0552 mL | 10.2762 mL | 20.5524 mL | 51.3811 mL |
| 5 mM | 0.4110 mL | 2.0552 mL | 4.1105 mL | 10.2762 mL | |
| 10 mM | 0.2055 mL | 1.0276 mL | 2.0552 mL | 5.1381 mL | |
| 15 mM | 0.1370 mL | 0.6851 mL | 1.3702 mL | 3.4254 mL | |
| 20 mM | 0.1028 mL | 0.5138 mL | 1.0276 mL | 2.5691 mL | |
| 25 mM | 0.0822 mL | 0.4110 mL | 0.8221 mL | 2.0552 mL | |
| 30 mM | 0.0685 mL | 0.3425 mL | 0.6851 mL | 1.7127 mL | |
| 40 mM | 0.0514 mL | 0.2569 mL | 0.5138 mL | 1.2845 mL | |
| 50 mM | 0.0411 mL | 0.2055 mL | 0.4110 mL | 1.0276 mL | |
| 60 mM | 0.0343 mL | 0.1713 mL | 0.3425 mL | 0.8564 mL | |
| 80 mM | 0.0257 mL | 0.1285 mL | 0.2569 mL | 0.6423 mL | |
| 100 mM | 0.0206 mL | 0.1028 mL | 0.2055 mL | 0.5138 mL |
Pitstop 2d Related Classifications
HY-186018 Related Classifications
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Keywords
- Pitstop 2d
- Clathrin
- VSV
- clathrin inhibitor
- clathrin terminal domain
- clathrin-mediated endocytosis
- CME
- endocytosis inhibitor
- Pitstop 2 derivative
- epsin
- SNX9
- transferrin uptake
- VSV entry
- vesicular stomatitis virus
- low cytotoxicity
- nuclear permeability barrier
- Cos7 cells
- HeLa cells
- clathrin-coated pits
- protein-protein interaction inhibitor
- Inhibitor
- inhibitor
- inhibit