Pitstop 2

Catalog No.S9670 Batch:S967001

Technical Data

Formula

C₂₀H₁₃BrN₂O₃S₂

Molecular Weight

473.36

CAS No.

1332879-52-3

Solubility (25°C)*

In vitro

DMSO

95 mg/mL (200.69 mM)

Water

Insoluble

Ethanol

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	4.75mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 95 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil	0.8mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 16 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

Pitstop 2 is an inhibitor of the interaction of amphiphysin with the amino terminal domain of clathrin that inhibits both clathrin-dependent endocytosis (CDE) and clathrin independent endocytosis (CIE).

Targets

CIE ^[1]

In vitro

Pitstop 2 is an inhibitor of the interaction of amphiphysin with the amino terminal domain of clathrin and inhibits clathrin-dependent endocytosis (CDE) and clathrin independent endocytosis (CIE) in cells. The effects of pitstop 2 are not restricted to inhibition of clathrin since knockdown of clathrin fails to rescue the inhibition of endocytosis of CIE proteins by the drug. Thus pitstop 2 has additional cellular targets besides the amino terminal domain of clathrin and thus cannot be used to distinguish CIE from CDE.^[1]

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines Hela cells Concentrations 20 μM Incubation Time 15 min Method Hela cells are preincubated with DMSO (control) or 20 μM pitstop 2 or pitstop 2-negative control for 15 min. Cells are allowed to internalize Alexa594-Transferrin and antibodies to MHCI for 30 min at 37℃ in the presence of DMSO or drugs. Surface bound antibodies are removed by low pH wash to visualize the internalized proteins (Internal) prior to fixation or cells are fixed immediately after the internalization to label for the internal and surface pools (Total) of the protein. Cells are then labeled with secondary antibodies to detect MHCI.

Cell Assay:^{^[1]}

Cell lines
Hela cells
Concentrations
20 μM
Incubation Time
15 min
Method
Hela cells are preincubated with DMSO (control) or 20 μM pitstop 2 or pitstop 2-negative control for 15 min. Cells are allowed to internalize Alexa594-Transferrin and antibodies to MHCI for 30 min at 37℃ in the presence of DMSO or drugs. Surface bound antibodies are removed by low pH wash to visualize the internalized proteins (Internal) prior to fixation or cells are fixed immediately after the internalization to label for the internal and surface pools (Total) of the protein. Cells are then labeled with secondary antibodies to detect MHCI.

References

[1] Dipannita Dutta, et al. PLoS One. 2012;7(9):e45799.

Selleck's Pitstop 2 has been cited by 6 publications

The autotaxin-LPA axis promotes membrane trafficking and secretion in yolk sac visceral endoderm cells [ Biol Open, 2023, 10.1242/bio.060081]	PubMed: 37795611
Baricitinib attenuates the proinflammatory phase of COVID-19 driven by lung-infiltrating monocytes [ Cell Rep, 2022, 39(11):110945]	PubMed: 35688145
The D614G mutation redirects SARS-CoV-2 spike to lysosomes and suppresses deleterious traits of the furin cleavage site insertion mutation [ Sci Adv, 2022, 8(51):eade5085]	PubMed: 36563151
TRESK channel contributes to depolarization-induced shunting inhibition and modulates epileptic seizures [ Cell Rep, 2021, 36(3):109404]	PubMed: 34289346
VPS34 regulates dynamin to determine the endocytosis of mitochondria-targeted zinc oxide nanoparticles in human osteosarcoma cells [ J Mater Chem B, 2021, 9(11):2641-2655]	PubMed: 33683276
Arginine-Arginine-Leucine Peptide Targeting Heat Shock Protein 70 for Cancer Imaging [ Mol Pharm, 2021, 10.1021/acs.molpharmaceut.1c00273]	PubMed: 34491767

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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