Bisindolylmaleimide IX (Ro 31-8220) Mesylate

Catalog No.S7207 Batch:S720702

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Technical Data

Formula

C25H23N5O2S.CH4O3S
Molecular Weight 553.65 CAS No. 138489-18-6
Solubility (25°C)* In vitro DMSO 100 mg/mL (180.61 mM)
Water Insoluble
Ethanol Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5% DMSO 95% Corn oil
0.41mg/ml Taking the 1 mL working solution as an example, add 50 μL of 8.2 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. 
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
5.0mg/ml Taking the 1 mL working solution as an example, add 50 μL of 100 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. 
* <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 Bisindolylmaleimide IX (Ro 31-8220 Mesylate) is a pan-PKC inhibitor with IC50 of 5 nM, 24 nM, 14 nM, 27 nM, and 24 nM for PKC-α, PKC-βI, PKC-βII, PKC-γ, and PKC-ε, respectively, and also shows potent inhibition against MAPKAP-K1b, MSK1, GSK3β and S6K1.
Targets
PKCα [1]
(Cell-free assay)		 PKCβ2 [1]
(Cell-free assay)		 PKCβ1 [1]
(Cell-free assay)		 PKCε [1]
(Cell-free assay)		 PKCγ [1]
(Cell-free assay)
5 nM 14 nM 24 nM 24 nM 27 nM
In vitro Ro 31-8220 inhibits rat brain PKC activity with IC50 of 23 nM, and does not show any high degree of selectivity between PKC-α, PKC-β, PKC-γ, and PKC-ε. [1] Ro 31-8220 also inhibits MSK1, MAPKAPK1, RSK, GSK3β and S6K1 with a potency similar to that for PKC. In addition, Ro 31-8220 inhibits voltage-dependent Na+ channels. [2] Ro 31-8220 alters cellular protein kinase C localization and potently inhibits growth of A549 and MCF-7 cells with IC50 of 0.78 μM and 0.897 μM, respectively. [3] RO 31-8220 enhances epinephrine-induced platelet aggregation in catecholamine hypo-responsive platelets by enhancing Akt phosphorylation. [4] Ro 31-8220 significantly decreases apoE secretion from primary human macrophages by inhibiting vesicular transport of apoE to the plasma membrane without significantly affecting apoE mRNA or apoE protein levels. [5]
In vivo In MLP−/− Mice, Ro 31-8220 (6 mg/kg/d, s.c.) results in a significant increase in cardiac contractility. [6]

Protocol (from reference)

Kinase Assay:

[1]
  • Assay of PKC Activity

    Assay mixtures contain 0.2 mg/mL peptide-gamma, 10 μM MgCl2, 0.6 mM CaCl2, 10 μM [γ-32P]ATP, 1.25 mg/mL phosphatidylserine and 1.25 ng/mL phorbol 12-myristate 13-acetate in 20 mM Hepes (pH 7.5), 2 mM EDTA, 1 mM dithiothreitol and 0.02% (w/v) Triton X-100. Peptide-γ is a synthetic peptide, GPRPLFCRKGSLRQKW, resembling the PKC-γ pseudosubstrate site, except that a serine residue replaces the pseudosubstrate alanine, converting the peptide from an inhibitor into a substrat. The assays are started by the addition of 2.5 m-units of enzyme, incubated at 30 °C for 10 min and terminated by spotting on to P81 paper, followed by extensive washing in 75 mM orthophosphoric acid. The papers are then washed in ethanol, dried, and incorporated radioactivity is determined by liquidscintillation spectroscopy.
Cell Assay:

[3]
  • Cell lines

    Human A549 lung and MCF-7 breast carcinoma cells

  • Concentrations

    2.5 μM

  • Incubation Time

    24-48 hours

  • Method

    Human A549 lung and MCF-7 breast carcinoma cells are obtained from the European Collection of Animal Cell Cultures. Cells (passage number 10-30) are cultured in an atmosphere of 5% carbon dioxide, the former in Ham's F-12 medium with penicillin/streptomycin, the latter in minimum essential medium (Eagle's modification) with additional pyruvate (1 mM) and non-essential amino acids. Both media are supplemented with 10% FCS and glutamine (2 mM). Cells are subcultured routinely twice weekly to maintain logarithmic growth. For cell proliferation studies cells are seeded and incubated with 3 ml of medium including agents, which is replenished at intervals of 48 h (A549) or 72 h (MCF-7). Following incubation for 4 days (A549) or 6 days (MCF-7) with drugs, cell number is assessed using a Coulter Counter Model ZM. In order to achieve PKC depletion, cells are incubated for 24 h with bryostatin 1 (1 μM). Under these conditions growth inhibition caused by bryostatin 1 is negligible. Bryostatin is removed by extensive washing of the cells followed by a 2 h recovery period. In previous work using the A549 cell line this washing procedure has been shown to eliminate bryostatin-mediated effects. The cells are then incubated for a further 24 h with staurosporine, RO 31-8220, UCN-01 or H-7. In some experiments cells are incubated with inhibitor for 48 rather than 24 h, in this case bryostation was not removed and left in the incubate. After removal of agents inhibition of DNA synthesis is evaluated by measurement of [3H]Tdr incorporation into cell. Radioactivity is counted using a Packard 1500 Tricarb scintillation counter.
Animal Study:

[6]
  • Animal Models

    MLP−/− Mice

  • Dosages

    6 mg/kg/d

  • Administration

    s.c.

Selleck's Bisindolylmaleimide IX (Ro 31-8220) Mesylate has been cited by 27 publications

Alternative splicing of HDAC7 regulates its interaction with 14-3-3 proteins to alter histone marks and target gene expression [ Cell Rep, 2023, 42(3):112273] PubMed: 36933216
Systems immunology-based drug repurposing framework to target inflammation in atherosclerosis [ Nat Cardiovasc Res, 2023, 2(6):550-571] PubMed: 37771373
Alternative splicing of apoptosis genes promotes human T cell survival [ Elife, 2022, 11e80953] PubMed: 36264057
SS18-SSX drives CREB activation in synovial sarcoma [ Cell Oncol (Dordr), 2022, 10.1007/s13402-022-00673-w] PubMed: 35556229
Establishment and characterization of immortalized sweat gland myoepithelial cells [ Sci Rep, 2022, 12(1):7] PubMed: 34997030
TaurineAttenuates the Hypotaurine-Induced Progression of CRCviaERK/RSK Signaling [ Front. Cell Dev. Biol, 2021-, Volume 9] PubMed: None
Reversible phosphorylation of cyclin T1 promotes assembly and stability of P-TEFb [ Elife, 2021, 10e68473] PubMed: 34821217
Taurine Attenuates the Hypotaurine-Induced Progression of CRC via ERK/RSK Signaling [ Front Cell Dev Biol, 2021, 9:631163] PubMed: 33937232
Taurine Attenuates the Hypotaurine-Induced Progression of CRC via ERK/RSK Signaling [ Front Cell Dev Biol, 2021, 9:631163] PubMed: 33937232
Taurine Attenuates the Hypotaurine-Induced Progression of CRC via ERK/RSK Signaling [ Front Cell Dev Biol, 2021, 9:631163] PubMed: 33937232

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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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