Overview

  • Product namePitstop® 2
  • Description
    Novel, selective cell-permeable clathrin inhibitor
  • Biological descriptionNovel, selective, cell membrane permeable clathrin inhibitor. Competitively inhibits clathrin terminal domain to selectively inhibit clathrin mediated endocytosis (CME) (IC50 = 12 μM for inhibition of amphiphysin association of clathrin TD). Interferes with receptor mediated endocytosis (RME), entry of HIV and synaptic vesicle recycling.
  • Purity> 98%
  • General notesSold under exclusive licence from Children's Medical Research Institute and Newcastle Innovation Ltd. Pitstop® is a trademark of Freie Universitat Berlin, Newcastle Innovation Ltd. and Children's Medical Research Institute

Properties

    Pitstop® 2 images

    • A) Pitstop® 2 reversibly inhibits Tf uptake. After 15 min preincubation HeLa cells were incubated with Alexa Fluor® 568-Tf in the presence of DMSO or 30 µM Pitstop 2 for 15 min. Tf uptake is seen to resume after washout of the drug for 1 hr. Scale bar, 10 mm. B) Reversibility and dose dependence of Pitstop® 2-mediated inhibition of Tf uptake. Data represent SEM (n = 3 independent experiments; *p < 0.05, ***p < 0.0001). C) Pitstop® 2 inhibits EGF uptake. HeLa cells pretreated with 30 µM pitstop 2 or DMSO for 15 min were incubated for 15 min with Alexa Fluor® 488-EGF in the continued presence of inhibitor. Data represent SEM (n = 3 independent experiments; ***p < 0.0001). D) Pitstop 2 does not interfere with AP-2-mediated cargo sequestration into CCPs. TIRF microscopy images of Cos7 cells pretreated with DMSO or 30 µM Pitstop 2 for 15 min were incubated with Alexa Fluor® 488-EGF at 8oC in the continued presence of inhibitor and immunostained for AP-2a (red). Scale bar, 4 mm. E) Pearson’s correlation between Alexa Fluor® 488-EGF and AP-2. Data represent SEM (n = 3 independent experiments; *p < 0.05).††This material was published for Cell, 146(3):471-484. von Kleist et al., Role of the Clathrin Terminal Domain in Regulating Coated Pit Dynamics Revealed by Small Molecule Inhibition. Copyright Elsevier (2011), with permission from Elsevier. Alexa Fluor® is a registered trademark of Life Technologies. Alexa Fluor® dye conjugates contain(s) technology licensed to Abcam by Life Technologies.

    Protocols

    References for Pitstop® 2 (ab120687)

    This product has been referenced in:
    • von Kleist L  et al. Role of the clathrin terminal domain in regulating coated pit dynamics revealed by small molecule inhibition. Cell 146:471-84 (2011). Read more (PubMed: 21816279) »

    See all 27 Publications for this product

    Product Wall

    As stated on page 7 of our protocol booklet for ab120687, longer incubation times (such as greater than 30 minutes) may lead to non-specific effects and are therefore not recommended.



    To answer this question I think its important to highlight the following note found on page 4-5 of the protocol booklet:

    -The final working concentration of DMSO should generally be between 0.3 and 1%. Low concentrations of DMSO (i.e. 0.1%) ma...

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    Thank you for your inquiry.

    I heard back from the lab and our chemists believe that this should have had no effect on the stability of the compound. Please tell the customer that the compound should be fine to use.
    We will remove the &ls...

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    We recommend use of serum free media (containing 10 mM HEPES; pH 7.4). Up to 0.1% serum can be tolerated for some cell lines, however this has not been extensively investigated. Longer incubation times may lead to non-specific effects and is therefor...

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    Unfortunately as this product is quite novel we were unable to find any specific citation of this product being used in Ovarian cancer cells. And as such, we were unable to find an ID50 value.

    However, we have a protocol book of guideline...

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    We cannot find any reason why the customers cells should be dying (we have not received any other reports of the compound being toxic to cells). Previous publications have only looked at HeLa and COS-7 cells ranging in concentrations between 20 - 30 uM...

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    The chemist here thinks that the white puff you are seeing is simply the DMSO meeting the PBS, and suggests that you will see the same puff when you pipet just DMSO, without any Pitstop 2™ dissolved in it. You might try that, as an easy check. He doubt...

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    Thank you for contacting us.

    Here are some guidelines for reconstituting and storing ab120687:

    A stock solution should be prepared by dissolving in 100% DMSO (fresh, sterile) to give a final concentration of 30 mM.
    Vortexing shoul...

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    Thank you for contacting us. The product ab120687 is not available reconstituted in DMSO, I am sorry to report. Here are some guidelines for reconstituting and storing it:

    A stock solution should be prepared by dissolving in 100% DMSO (fres...

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    Thank you for contacting us.

    As we do not undertake our own research we are only able to provide information available in the current literature. Von Kleist et al., (2011) found no cytotoxicity in HeLa cells after 8, 20 or 72 hours and concl...

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    1-10 of 13 Abreviews or Q&A

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE, NOT FOR USE IN HUMANS"