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    InvivoChem Cat #: V1674
    CAS #: 289905-88-0Purity ≥98%

    Description: TRAM-34 (TRAM34; TRAM 34 Triarylmethane-34) is an inhibitor of Ca2+-activated K+ channel (IKCa1, KCa3.1) with important biological activity. It inhibits Ca2+-activated K+ channel with a Kd of 20 nM, and exhibits 200- to 1500-fold higher selectivity over cloasely related ion channels.. 

    References: Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6; Circulation. 2003 Sep 2;108(9):1119-25.

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    Molecular Weight (MW)344.84
    CAS No.289905-88-0
    Storage-20℃ for 3 years in powder form
    -80℃ for 2 years in solvent
    Solubility (In vitro)DMSO: 0.4 mg/mL (1.15 mM)
    Water: <1 mg/mL
    Ethanol: <1 mg/mL
    Solubility (In vivo)30% PEG400+0.5% Tween80+5% Propylene glycol: 30mg/mL

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    In Vitro

    In vitro activity: Unlike clotrimazole, TRAM-34 selectively inhibits IKCa1 without blocking cytochrome P450 enzyme (CYP3A4). TRAM-34 potently inhibits cloned IKCa1 channel in IKCa1-transfected COS-7 cells as well as native IKCa currents in human T lymphocytes and T84 cells with Kd of 20 nM, 25 nM, and 22 nM, respectively, more potently than clotrimazole with Kd of 70 nM, 100 nM, and 90 nM, respectively. TRAM-34 exhibits 200- to 1,500-fold selectivity over other ion channels such as KV, BKCa, SKCa, Na+, CRAC and Cl- channels. TRAM-34 significantly inhibits anti-CD3 Ab or PKC-activator PMA plus calcium-ionophore ionomycin induced activation of human T lymphocytes with IC50 of 295-910 nM and 85-830 nM, respectively. TRAM-34 (5 μM) does not inhibit cell viability of human T lymphocytes or several cell lines. TRAM-34 significantly inhibits EGF-induced IKCa1 up-regulation, and EGF-stimulated proliferation of A7r5 cells with IC50 of 8 nM. TRAM-34 treatment inhibits proliferation of human endometrial cancer (EC) cells, and blocks EC cell cycle at G0/G1 phase. Inhibition of the IKCa1 channel by TRAM-34 (1-30 μM) leads to dose-dependent suppression of the proliferation but not apoptosis of LNCaP and PC-3 prostate cancer (PCa) cells, involving an increase of p21Cip1 and cell arrest in the G1 cycle

    Kinase Assay: The human IKCa1 is cloned and expressed in COS-7 cells. Cells are studied in the whole-cell configuration of the patch-clamp technique. The holding potential is 280 mV. The internal pipette solution contains: 145 mM K+ aspartate, 2 mM MgCl2, 10 mM Hepes, 10 mM K2EGTA, and 8.5 mM CaCl2 (1 μM free Ca2+), pH 7.2, 290-310 mOsm. To reduce currents from the native chloride channels in COS-7 cells, Na+ aspartate Ringer is used as an external solution: 160 mM Na+ aspartatey/4.5 mM KCl/2 mM CaCl2/1 mM MgCl2/5 mM Hepes, pH 7.4/290-310 mOsm. IKCa currents in COS-7 cells are elicited by 200-ms voltage ramps from -120 mV to 40 mV applied every 10 seconds and the reduction of slope conductance at -80 mV by TRAM-34 taken as a measure of channel block.

    Cell Assay: Cells (Human T lymphocytes, Jurkat E6-1, MEL, C2F3, CHO, COS-7, L929, NGP, NLF, and RBL-2H3) are exposed to TRAM-34 for 48 hours. After 48 hours, cells are harvested by suction (suspension cells) or by trypsinization (adherent cell lines), centrifuged, resuspended in 0.5 mL PBS containing 1 μg/mL propidium iodide (PI), and red fluorescence measured on a FACScan flow cytometer. The percentage of dead cells is determined by their PI uptake, 104 cells of every sample being analyzed.

    In VivoTRAM-34 treatment at ~500-1,000 times the channel-blocking dose (0.5 mg/kg/day) for 7 days is nontoxic to mice. Administration of TRAM-34 at 120 mg/kg/day significantly reduces intimal hyperplasia by ~40% in a rat model of balloon catheter injury (BCI). Consistent with its in vitro role in inhibiting the proliferation of EC cells, TRAM-34 treatment at 30 μM slows the development of HEC-1-A tumor in vivo.
    Animal modelSprague-Dawley rats subjected to BCI of the left CA by use of a 2F Fogarty embolectomy catheter
    Formulation & DosageFormulated in peanut oil; 120 mg/kg/day; s.c. injection 

    Br J Pharmacol. 2001 May;133(1):193-9; Horm Metab Res. 1996 Sep;28(9):469-87; Indian J Exp Biol. 2009 Oct;47(10):804-10.

    These protocols are for reference only. InvivoChem does not independently validate these methods.


    (A) IKCa1 currents in COS-7 cells blocked by TRAM-34. (B) Dose response for IKCa1 channel block by TRAM-34. Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.
    Effect of TRAM-34 on anti-CD3 Ab- (A) or PMA + ionomycin- (B) stimulated [(3H)-TdR incorporation by resting and preactivated lymphocytes. Proc Natl Acad Sci U S A. 2000 Jul 5;97(14):8151-6.


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