Pitstop 2c
Based on 1 Customer Validation
Pitstop 2c is a clathrin terminal domain inhibitor with an IC50 of 1.8 μM. Pitstop 2c blocks the binding of epsin 1 to the clathrin terminal domain and impairs epsin 1-mediated membrane curvature formation at the early stage of clathrin-mediated endocytosis (CME). Pitstop 2c enhances the clathrin-independent endocytosis of galectin-3 and alters its intracellular distribution. Pitstop 2c causes mild defects in lysosomal acidification, as well as a slight increase in mitochondrial number but a reduction in average mitochondrial size. Pitstop 2c can be used in studies related to vesicular stomatitis virus infection.
Para uso exclusivo en investigación. No vendemos a pacientes.
- Pureza : 98.54%
- Fòrmula: C26H18N2O4S2
- Peso molecular:486.56
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Almacenamiento:
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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Actividad biológica
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Chemical Information
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Solvente y solubilidad
- Protocolo
- Pureza y Documentación
- Referencias
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Help & FAQs
Actividad biológica
Descripciòn
IC50 & Target
IC50: <2 μM (Clathrin TD)[1]
In Vitro
Pitstop 2c inhibits clathrin terminal domain-amphiphysin binding in vitro with an IC50 of 1.8 μM[1].
Pitstop 2c (25-100 μM; 8-20 h) exhibits low cytotoxicity in HEK293 cells[1].
Pitstop 2c (25-100 μM; 8-20 h) exhibits low cytotoxicity in Cos7 cells[1].
Pitstop 2c (30 μM; 15 min) does not disrupt the nuclear permeability barrier in HeLa cells[1].
Pitstop 2c (30 μM; 20 min) inhibits transferrin CME in HeLa cells with an IC50 of 13 μM, and nearly completely blocks uptake[1].
Pitstop 2c (45 μM) reversibly inhibits transferrin CME in HEK293T cells[1].
Pitstop 2c (45 μM) reversibly inhibits transferrin CME in Cos7 cells[1].
Pitstop 2c (low, non-toxic dose; 15 min pre-treatment, 5 min imaging) perturbs CCP dynamics in Cos7 cells stably expressing eGFP-clathrin light chain α, reducing the fraction of dynamic coated pits[1].
Pitstop 2c (30 μM; 20 min) impairs clathrin dynamics in the perinuclear region of Cos7 cells stably expressing eGFP-clathrin light chain α, reducing maximal fluorescence recovery to 44.4%[1].
Pitstop 2c (45 μM; 15 min) does not impair clathrin-independent fluid-phase endocytosis of dextran in HeLa cells[1].
Pitstop 2c (15 min pre-incubation, 2 min galectin-3 internalization) enhances clathrin-independent endocytosis of galectin-3 and alters its subcellular distribution in HeLa cells[1].
Pitstop 2c (100 μM; 1 h) displaces Epsin 1 from clathrin and increases clathrin association with SNX9 in Cos7 cells stably expressing eGFP-clathrin light chain α[1].
Pitstop 2c (four-fold molar excess relative to clathrin terminal domain) binds to the clathrin box site of the clathrin terminal domain in a distinct U-shaped conformation, forming specific interactions with residues R64 and K96[1].
Pitstop 2c (20 μM; 15 min pre-incubation, 1 h viral infection) inhibits VSV entry into Vero-E6 cells by >60%[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:HEK293 cells
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Concentration:25 μM; 50 μM; 100 μM
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Incubation Time:8 h; 20 h
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Result:Caused minimal cytotoxicity at all tested concentrations and incubation times, with cytotoxicity percentages far lower than the reference compound Pitstop 2.
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Cell Line:Cos7 cells
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Concentration:25 μM; 50 μM; 100 μM
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Incubation Time:8 h; 20 h
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Result:Caused minimal cytotoxicity at all tested concentrations and incubation times, with cytotoxicity percentages far lower than the reference compound Pitstop 2.
Chemical Information
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Appearance Solid
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Peso molecular 486.56
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Fòrmula C26H18N2O4S2
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Color Light yellow to yellow
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SMILES
O=S(C1=C2C=CC=CC2=CC=C1)(NC(S/C3=C\C4=C(C=CC=C4)OC5=CC=CC=C5)=NC3=O)=O
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Envío
Room temperature in continental US; may vary elsewhere.
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Almacenamiento
4°C, sealed storage, away from moisture
* In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture)
Solvente y solubilidad
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.
Protocolo
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Research Protocol for Infectious Diseases
Infectious-disease experiments test how pathogens interact with host barriers, innate immune receptors, inflammatory signaling, pathogen replication, and tissue injury; pattern-recognition receptors such as TLRs, RIG-I-like receptors, NOD-like receptors, and inflammasomes detect microbial molecules and activate NF-κB, interferon, and cytokine responses. The central hypothesis is that infection severity reflects the balance between pathogen burden and host response: protective inflammation restricts pathogen growth, whereas excessive or mislocalized inflammation contributes to tissue damage and disease phenotype. Unresolved questions include which host pathways are protective versus pathogenic, why some infection models fail to translate to human disease, and which combined readouts best predict clinically relevant infection outcomes.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Lysosome and acidic-vesicle live-cell staining
Lysosome and acidic-vesicle live-cell staining detects acidic intracellular compartments by using membrane-permeant acidotropic probes that accumulate in low-pH vesicles, including lysosomes, late endosomes, autolysosomes, and acidic phagosomes. LysoTracker staining is commonly used as an intensity-based readout of acidic lysosomal compartment abundance or enlargement, while acridine orange produces green fluorescence in less concentrated compartments and red fluorescence after concentration-dependent accumulation in acidic vesicular organelles. Loss or reduction of acridine-orange red signal can be used as a readout of lysosomal membrane permeabilization or reduced acidic-vesicle integrity. This protocol is designed for live cultured cells and can be adapted for fluorescence microscopy, high-content imaging, plate-reader readout, or flow cytometry when the selected literature supports the readout. Because these dyes report acidotropic accumulation rather than lysosome identity alone,
Pureza y Documentación
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Ficha de datos (282 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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Instrucciones de manejo (2659 KB)
Referencias
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 |
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| 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 2c Related Classifications
HY-186017 Related Classifications
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