1. TGF-beta/Smad
  2. TGF-β Receptor

RepSox (Synonyms: E-616452; SJN 2511)

Cat. No.: HY-13012 Purity: 99.94%
Handling Instructions

RepSox is a potent and selective of the TGFβR-1/ALK5 inhibitor, inhibits ALK5 autophosphorylation with IC50 of 4 nM.

For research use only. We do not sell to patients.

RepSox Chemical Structure

RepSox Chemical Structure

CAS No. : 446859-33-2

Size Price Stock Quantity
Free Sample (0.5-1 mg)   Apply now  
10 mM * 1 mL in DMSO USD 79 In-stock
Stock in the United States
Estimated Time of Arrival: December 31
5 mg USD 72 In-stock
Stock in the United States
Estimated Time of Arrival: December 31
10 mg USD 108 In-stock
Stock in Sweden
Estimated Time of Arrival: December 31
50 mg USD 420 In-stock
Stock in Sweden
Estimated Time of Arrival: December 31
100 mg   Get quote  
200 mg   Get quote  

* Please select Quantity before adding items.

    RepSox purchased from MCE. Usage Cited in: Oncotarget. 2017 Jun 19;8(36):60159-60172.

    SH-SY5Y (A) and HEK293 cells (B) are treated with 25 μM RepSox (Rep) in absence or presence of dPA or dPPA for 72 h, and the Western blotting results show that dPA or dPPA increases the expression in the presence of RepSox. RepSox does not cause significant alteration of furin protein compared to control. The expression of furin remains significantly increased after dPA or dPPA in the presence of RepSox.
    • Biological Activity

    • Protocol

    • Technical Information

    • Purity & Documentation

    • References

    Description

    RepSox is a potent and selective of the TGFβR-1/ALK5 inhibitor, inhibits ALK5 autophosphorylation with IC50 of 4 nM.

    IC50 & Target

    IC50: 4 nM (ALK5 autophosphorylation)[1]

    In Vitro

    RepSox also inhibits ATP binding to ALK5 with IC50 of 23 nM. RepSox shows potent activity in both binding and cellular assays and exhibits selectivity over p38 mitogen-activated protein kinase. with IC50 of >16 μM[1]. RepSox act s as an inhibitor of the Tgfβ1 kinase. Treatment with 25 μM RepSox almost completely eliminates Smad3 phosphorylation, indicating that RepSox strongly inhibits Tgfβ signaling in somatic cells. RepSox is most effective at replacing Sox2 during days 10-11 after transduction and that therefore cultures of Oct4, Klf4, and cMyc-transduced MEFs give rise to intermediates capable of responding to RepSox treatment. These intermediates appear at day 4 post-transduction and peak at days 10-11. Treatment with RepSox decreased the proportion of cells in G2/M phase of the cell cycle, indicating it does not increase the proliferation rate of these partially reprogrammed cells[2].

    References
    Preparing Stock Solutions
    Concentration Volume Mass 1 mg 5 mg 10 mg
    1 mM 3.4804 mL 17.4022 mL 34.8044 mL
    5 mM 0.6961 mL 3.4804 mL 6.9609 mL
    10 mM 0.3480 mL 1.7402 mL 3.4804 mL
    Please refer to the solubility information to select the appropriate solvent.
    Kinase Assay
    [1]

    The kinase domain of ALK5 is cloned by PCR and expressed in a baculovirus/Sf9 cells system. The protein is 6-His tagged in the C terminus and purified by affinity chromatography using a Ni2+column, and the obtained material is used to assess compound activity in an autophosphorylation assay. Purified enzyme (10 nM) is incubated in 50 μL of Tris buffer (Tris 50 mM, pH 7.4; NaCl, 100 mM; MgCl2, 5 mM; MnCl2, 5 mM; and DTT, 10 mM). The enzyme is preincubated with different concentrations of RepSox (0.1% DMSO final concentration in the test) for 10 min at 37°C. The reaction is then initiated by the addition of 3 μM ATP (0.5 μCi γ-33P-ATP). After 15 min at 37°C, phosphorylation is stopped by the addition of SDS−PAGE sample buffer (50 mM Tris-HCl, pH 6.9, 2.5% glycerol, 1% SDS, and 5% β-mercaptoethanol). The samples are boiled for 5 min at 95°C and run on a 12% SDS−PAGE. Dried gels are exposed to a phosphor screen overnight. ALK5 autophosphorylation is quantified using a Storm imaging system[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    Cell Assay
    [1]

    RepSox is dissolved in DMSO and stored, and then diluted with appropriate media (DMSO 1%) before use[1].

    To test anti-TGF-β activity of compounds, HepG2 cells are seeded in 96 well microplates at a concentration of 35000 cells per well in 200 μL of serum-containing medium. The microplates are then placed for 24 h in a cell incubator at 37°C, 5% CO2 atm. RepSox dissolved in DMSO are then added at concentrations of 50 nM to 10 μM (final concentration of DMSO 1%) for 30 min prior to the addition of recombinant TGF-β (1 ng/mL). After an overnight incubation, the cells are washed with PBS and lysed by addition of 10 μL of passive lysis buffer. Inhibition of luciferase activity relative to control groups is used as a measure of compound activity. A concentration−response curve is constructed from which an IC50 value is determined graphically[1]. MCE has not independently confirmed the accuracy of these methods. They are for reference only.

    References
    Molecular Weight

    287.32

    Formula

    C₁₇H₁₃N₅

    CAS No.

    446859-33-2

    SMILES

    CC1=NC(C2=NNC=C2C3=NC4=CC=CN=C4C=C3)=CC=C1

    Storage
    Powder -20°C 3 years
      4°C 2 years
    In solvent -80°C 6 months
      -20°C 1 month
    Shipping

    Room temperature in continental US; may vary elsewhere

    Solvent & Solubility

    DMSO: ≥ 52 mg/mL

    * "<1 mg/mL" means slightly soluble or insoluble. "≥" means soluble, but saturation unknown.

    Inquiry Online

    Your information is safe with us. * Required Fields.

    Product name

     

    Salutation

    Applicant name *

     

    Email address *

    Phone number *

     

    Organization name *

    Country or Region *

     

    Requested quantity *

    Remarks

    Bulk Inquiry

    Inquiry Information

    Product Name:
    RepSox
    Cat. No.:
    HY-13012
    Quantity: