Ruxolitinib (INCB018424)

Catalog No.S1378

Ruxolitinib (INCB018424) Chemical Structure

Molecular Weight(MW): 306.37

Ruxolitinib (INCB018424) is the first potent, selective, JAK1/2 inhibitor to enter the clinic with IC50 of 3.3 nM/2.8 nM in cell-free assays, >130-fold selectivity for JAK1/2 versus JAK3.

Size Price Stock Quantity  
In DMSO USD 208 In stock
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Cited by 34 Publications

4 Customer Reviews

  • STAT3 phosphorylation as determined by phospho flow, mixed lymphocyte reactions containing BALB/c spleen-derived CD4+ T cells co-cultured with or without C57BL/6 BM-derived DC preactivated with 20 ng/mL LPS.

    Blood 2014 123(24), 3832-42. Ruxolitinib (INCB018424) purchased from Selleck.

    BMDMs were isolated from wild-type mice and incubated in the different concentrations of Ruxolitinib for 1 h before stimulation with 500 U/ml IFN-β for 30 min. Levels of GAPDH as well as total and phospho-Tyr705 STAT3 were determined by immunoblotting.

    J Immunol 2012 189(6), 2784-92. Ruxolitinib (INCB018424) purchased from Selleck.

  • Miniscule 10 PFU amount of VA7-EGFP results in productive infection only in CT26LacZ cells whereas the self-limiting infection can be rescued with JAK/STAT inhibitor Ruxolitinib in CT26WT cells counteracting also the effects of exogenous IFNβ (100 U/ml) pre-treatment. Scale bar: 200 um.

    Gene Ther 2014 10.1038/gt.2014.83. Ruxolitinib (INCB018424) purchased from Selleck.

     

    HS578T cells were treated with indicated amount of inhibitor for 18 hr. The cells were lysed by cell lysis buffer (20 mM Tris-Cl (pH 8.0); 0.5 M NaCl; 0.25% Triton X-100; 1 mM EDTA; 1 mM EGTA; 10 mM β-glycerophosphate; 10 mM NaF; 300 µM Na3VO4; 1 mM benzamidine) containing 1 mM DTT and 2 µM PMSF. Western blot analyses were performed using cleared cell lysates resolved on sodium dodecyl sulfate (SDS)-polyacrylamide gels, transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore, Billerica, MA), and probed with specific antibodies using standard procedures. Chemiluminescence reagent was purchased from  Thermo Scientific (Rockford, IL). Horseradish peroxidase-conjugated secondary antibodies from Sigma (St. Louis, MO).

    Yong Weon Yi Georgetown University. Ruxolitinib (INCB018424) purchased from Selleck.

Purity & Quality Control

Choose Selective JAK Inhibitors

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Notes:

2. For more details, such as half maximal inhibitory concentrations (IC50s) and working concentrations of each inhibitor, please click on the link of the inhibitor of interest.
3. "+" indicates inhibitory effect. Increased inhibition is marked by a higher "+" designation.
4. Orange "√" refers to compounds which do inhibitory effects on the related isoform, but without specific value.

Biological Activity

Description Ruxolitinib (INCB018424) is the first potent, selective, JAK1/2 inhibitor to enter the clinic with IC50 of 3.3 nM/2.8 nM in cell-free assays, >130-fold selectivity for JAK1/2 versus JAK3.
Targets
JAK2 [1]
(Cell-free assay)
JAK1 [1]
(Cell-free assay)
2.8 nM 3.3 nM
In vitro

INCB018424 potently and selectively inhibits JAK2V617F-mediated signaling and proliferation in Ba/F3 cells and HEL cells. INCB018424 markedly increases apoptosis in a dose dependent manner in Ba/F3 cells. INCB018424 (64 nM) results in a doubling of cells with depolarized mitochondria in Ba/F3 cells. INCB018424 inhibits proliferating of erythroid progenitors from normal donors and polycythemia vera patients with IC50 of 407 nM and 223 nM, respectively. INCB018424 demonstrates remarkable potency against erythroid colony formation with IC50 of 67nM. [1]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
TF1 MV3LbY5ie2ViQYPzZZk> NFvTNZQzOCCvaX6= Mn\rSG1UVw>? MYHJcohq[mm2aX;uJI9nKEqDS{KgbY4hcHWvYX6gWGYyKGOnbHzzJIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gSXBQNWmwZIXj[YQhW1SDVEWgdIhwe3Cqb4L5cIF1cW:wIIfpeIghUUN3MDDv[kAxNjBzMt88US=> Mm[1NlI3QThyOES=
TF1 NYPGfZZuU2mwYYPlJGF{e2G7 MkH3NlAhdWmw NGLXTIFFVVOR MmTCTY5pcWKrdHnvckBw\iCMQVuxJIlvKGi3bXHuJHRHOSClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGlNPi2rbnT1Z4VlKFOWQWSzJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMkVOwG0> MljTNlI3QThyOES=
Human T cell MU\LbY5ie2ViQYPzZZk> MYLJcohq[mm2aX;uJI9nKEqDS{OvNUBqdiCqdX3hckBVKGOnbHzzJIV5eHKnc4PpcochS0R|IHHzd4V{e2WmIHHzJIlvcGmkaYTpc44hd2ZiSVyyMZN1cW23bHH0[YQhW1SDVEXhJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMkROwG0> M33XT|I{PTRyNkS4
Human monocyte NF;iSYxMcW6jc3WgRZN{[Xl? M1vofmlvcGmkaYTpc44hd2ZiSlHLNkBqdiCqdX3hckBud26xY4n0[ZMh\XiycnXzd4lv\yCFREG0JIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gS20uS1OILYP0bY12dGG2ZXSgV3RCXDWjIIDoc5NxcG:{eXzheIlwdiC5aYToJGlEPTBib3[gNE4xOjcQvF2= MkXvNlM2PDB4NEi=
Human monocyte MV;LbY5ie2ViQYPzZZk> M3G3RmlvcGmkaYTpc44hd2ZiSlHLNk8yKGmwIHj1cYFvKG2xbn;jfZRmeyCneIDy[ZN{cW6pIFPENVQh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBKTk6pYX3tZU1{fGmvdXzheIVlKFOWQWSxJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yM{JOwG0> NYDnTWp3OjN3NEC2OFg>
HEL M4[wb2N6fG:2b4jpZ{BCe3OjeR?= Ml75OUDPxE1? MX[0PEBp NITOd3REgXSxdH;4bYMhcW6mZYi9NVIvOiV? M2PkeFI2QTNzM{S5
SET-2 NXy0d5hnS3m2b4TvfIlkKEG|c3H5 NEHIdpk2KM7:TR?= MX20PEBp MXjDfZRwfG:6aXOgbY5l\Xh;MUiuO{U> NIr4[3ozPTl|MUO0PS=>
HT93A MU\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXqzNlAhdk1? Ml3rOUBl MmCySG1UVw>? MliwTY5pcWKrdHnvckBw\iCJQ2OtSkBqdmS3Y3XkJIdz[W63bH;jfZRq[yCmaX\m[ZJmdnSrYYTpc44> MVqyOVgxPTl4Mh?=
CMK M{W3PWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 Mkf3TY5pcWKrdHnvckBw\iCFTVugZ4FzenmrbnegeIhmKEqDS{PBOVczXiCvdYTheIlwdiClZXzsJJBzd2yrZnXyZZRqd25? NHPZcGYzPTN3MkGyOC=>
CMK NVLqdGtET3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MkLnTY5pcWKrdHnvckBw\iCFTVugZ4FzenmrbnegeIhmKEqDS{PBOlNFKG23dHH0bY9vKGOnbHygdJJwdGmoZYLheIlwdiC5aYToJGlEPTBib3[gNE4yPjNizszN MXSyOVM2OjF{NB?=
CMK NEnaVXRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NIDxRWlKdmirYnn0bY9vKG:oIFPNT{Bk[XK{eXnu[{B1cGViV2SgTmFMKGOnbHygdJJwdGmoZYLheIlwdiC5aYToJGlEPTBib3[gNE4xPzVizszN MWiyOVM2OjF{NB?=
NCI-H460 MmOyS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NIH3SIZFVVOR NXKzXGo{UUN3ME2wMlE{KM7:TR?= MkTiNlUzOTN4N{C=
NCI-H358 MlW0S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M4fCXmROW09? NGi0OJJKSzVyPUCuNUDPxE1? NEO0N|kzPTJzM{[3NC=>
A549 NUPk[mdmT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NUnrfYpoTE2VTx?= NGHIWVRKSzVyPUCuNFQh|ryP MWeyOVIyOzZ5MB?=
A549/DDP MUfHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M2jQ[2ROW09? M4fDN2lEPTB;MD6yNkDPxE1? NFPycpAzPTJzM{[3NC=>
NCI-H1299 NXrrRmJbT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MlHsSG1UVw>? Ml\6TWM2OD1yLkK4JO69VQ>? MmjyNlUzOTN4N{C=
NCI-H2347 MUHHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NX\zcGt7TE2VTx?= MorZTWM2OD1yLkG3JO69VQ>? NFLIS4IzPTJzM{[3NC=>
A549/DDP MnzrSpVv[3Srb36gRZN{[Xl? NYTCVXRJOzBibl2= MX20PEBp MoLkSG1UVw>? MknpSI94di2{ZXf1cIF1cW:wIH;mJHNVSVR|IIDoc5NxcG:{eXzheIlwdg>? MXSyOVIyOzZ5MB?=
NCI-H1299 NXLJU5RYTnWwY4Tpc44hSXO|YYm= NWXkcYJZOzBibl2= MkXtOFghcA>? NH3zVppFVVOR MVjEc5dvNXKnZ4XsZZRqd25ib3[gV3RCXDNicHjvd5Bpd3K7bHH0bY9v M2nuR|I2OjF|Nkew
NCI-H2347 NFfkPWdHfW6ldHnvckBCe3OjeR?= MlXNN|Ahdk1? NHHq[Fc1QCCq MWjEUXNQ NGLrN2tF\WO{ZXHz[UBqdiCEY3yyJIV5eHKnc4Ppc44> MUCyOVIyOzZ5MB?=
A549/DDP NFTCNmxCeG:ydH;zbZMhSXO|YYm= MVqzNEBvVQ>? M1P1c|Q5KGh? NWnCb|BHTE2VTx?= M2KxS2lv\HWldHnvckBw\iCjcH;weI9{cXN? NGLRTJEzPTJzM{[3NC=>
NCI-H1299 MUHBdI9xfG:|aYOgRZN{[Xl? MXuzNEBvVQ>? MnrQOFghcA>? NWC0V4lXTE2VTx?= MW\JcoR2[3Srb36gc4Yh[XCxcITvd4l{ NGPiOnMzPTJzM{[3NC=>
NCI-H2347 MVLBdI9xfG:|aYOgRZN{[Xl? NELJfWc{OCCwTR?= MkDiOFghcA>? NY[wVnJOTE2VTx?= NUPF[nQ3UW6mdXP0bY9vKG:oIHHwc5B1d3Orcx?= NWP2U2NFOjV{MUO2O|A>
Hep3B NUHkPJR5TnWwY4Tpc44hSXO|YYm= MWixJO69VQ>? MVKxOkBp MmrtSG1UVw>? M3zPZ2lueGGrcnXzJJRp\SClYYDhZ4l1gSCxZjDJTGNCNWG|c3;jbYF1\WRiZ4CxN|AhdXW2YX70d{B1dyCjY4TpeoUhW1SDVEOge4l1cCCLQ{WwJI9nKH53MDFOwG0> MXWyOFUxOTZ6OR?=
HepG2 NXHpTWVyTnWwY4Tpc44hSXO|YYm= NGHMdIkyKM7:TR?= NE\FW|UyPiCq NFrrbYdFVVOR MnHOTY1x[Wm{ZYOgeIhmKGOjcHHjbZR6KG:oIFnIR2Eu[XO|b3PpZZRm\CCpcEGzNEBufXSjboTzJJRwKHOrZ37hcEB1dyCVVFHUNy=> NIP2S2IzPDVyMU[4PS=>
Huh7 MnPHSpVv[3Srb36gRZN{[Xl? NXT2VGJ5OSEQvF2= NEfYPJIyPiCq Ml7ISG1UVw>? M{DuUmlueGGrcnXzJJRp\SClYYDhZ4l1gSCxZjDJTGNCNWG|c3;jbYF1\WRiZ4CxN|AhdXW2YX70d{B1dyC|aXfuZYwhfG9iU2TBWFM> NYi1VWFPOjR3MEG2PFk>
BaF3 NXWzV3FpU2mwYYPlJGF{e2G7 NX71fHpJQDBibl2= MVK2JIg> NWfGUmdMTE2VTx?= M2PmeXJm\HWlZYOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\sLiU2TBWFUhcW5iSlHLNnY3OTeILX31eIF1\WRiQlHGN{1GWE:UIHPlcIw> NWr2UVlzOjR{M{e3PVE>
DLD-1 NWHEV4txU2mwYYPlJGF{e2G7 MlPINlUh|ryP NV;lTHVrPDhiaB?= Mn3VSG1UVw>? NHTUT3lKdmirYnn0bY9vKG:oIFrBT|EheGixc4Doc5J6dGG2aX;u M4mzZlI1ODVyNUWw
RKO NV;HWI1YU2mwYYPlJGF{e2G7 MX2yOUDPxE1? NXLKSWhHPDhiaB?= MnHiSG1UVw>? MnixTY5pcWKrdHnvckBw\iCMQVuxJJBpd3OyaH;yfYxifGmxbh?= MoHmNlQxPTB3NUC=
DLD-1 M{i1XWtqdmG|ZTDBd5NigQ>? NHPVN3QzPSEQvF2= MV:0PEBp NYLMS3U4TE2VTx?= MXrJcohq[mm2aX;uJI9nKEqDS{KgdIhwe3Cqb4L5cIF1cW:w NVWyd4VnOjRyNUC1OVA>
RKO M33NbGtqdmG|ZTDBd5NigQ>? MYeyOUDPxE1? M{jYV|Q5KGh? NV[4S|lMTE2VTx?= NUDCNlZz\G:nczDuc5QhcW6qaXLpeEBLSUtzIIDoc5NxcG:{eXzheIlwdg>? M{\uUFI1ODVyNUWw
DLD-1 NXHsc5c6T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NXrsOXN1PTBizszN NG[xUYI1QCCq NFe2cZFFVVOR MV3JR|UxRTF3LkWxJO69VQ>? MoTNNlQxPTB3NUC=
RKO NWTlOmE1T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= M{jmWFUxKM7:TR?= MX[0PEBp NWX6d4RQTE2VTx?= NEf0e3BKSzVyPUG0Mlc3KM7:TR?= NV\ER|RyOjRyNUC1OVA>
DLD-1 MW\BdI9xfG:|aYOgRZN{[Xl? MUSyOUDPxE1? M{[zTVQ5KGh? NEHaVndFVVOR NFHXR4dKdmS3Y3XzJIFxd3C2b4Ppd{BjgSCjY4TpeoF1cW6pIHPhd5Bie2ViMx?= NHzVfZIzPDB3MEW1NC=>
RKO M1Kwc2Fxd3C2b4Ppd{BCe3OjeR?= MUCyOUDPxE1? M{PMUVQ5KGh? MV7EUXNQ MlT2TY5lfWOnczDhdI9xfG:|aYOgZpkh[WO2aY\heIlv\yClYYPwZZNmKDN? MkTzNlQxPTB3NUC=
HuH7 MVTHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NIjheZI2OCEQvF2= Ml3COFghcA>? M2rp[2ROW09? Mm\YQlgzLSC{ZXT1Z5Rqd25? MWWyN|k1OTh|Mh?=
SNU182 NIXMSG9Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M3HGWFUxKM7:TR?= NVnuZplCPDhiaB?= Mm\hSG1UVw>? MWC+OlQmKHKnZIXjeIlwdg>? MV[yN|k1OTh|Mh?=
SNU423 MVzHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? Mn7sOVAh|ryP MlvQOFghcA>? NV\XUWExTE2VTx?= MlX6QlgyLSC{ZXT1Z5Rqd25? MmL5NlM6PDF6M{K=
HuH7 MULGeY5kfGmxbjDBd5NigQ>? NGLycpg2OCEQvF2= Mki4NlQhcA>? MX\EUXNQ M{fOTGlvcGmkaYTpc44hd2ZiU2TBWFEh[W6mIGPURXQ{KHCqb4PwbI9zgWyjdHnvckB{cWewaX\pZ4FvfGy7 MVSyN|k1OTh|Mh?=
SNU182 NY\mSHo{TnWwY4Tpc44hSXO|YYm= NG[4ZpM2OCEQvF2= NY\sZ2RYOjRiaB?= MXLEUXNQ NHvBeG5KdmirYnn0bY9vKG:oIGPURXQyKGGwZDDTWGFVOyCyaH;zdIhwenmuYYTpc44he2mpbnnmbYNidnSueR?= MnPrNlM6PDF6M{K=
SNU423 NHSye3BHfW6ldHnvckBCe3OjeR?= MXW1NEDPxE1? M2fpW|I1KGh? NWKzToM1TE2VTx?= M4DhWmlvcGmkaYTpc44hd2ZiU2TBWFEh[W6mIGPURXQ{KHCqb4PwbI9zgWyjdHnvckB{cWewaX\pZ4FvfGy7 NWCzcoRJOjN7NEG4N|I>

... Click to View More Cell Line Experimental Data

In vivo INCB018424 (180 mg/kg, orally, twice a day) results in survive rate of greater than 90% by day 22 in a JAK2V617F-driven mouse model. INCB018424 (180 mg/kg, orally, twice a day) markedly reduces splenomegaly and circulating levels of inflammatory cytokines, and preferentially eliminated neoplastic cells, resulting in significantly prolonged survival without myelosuppressive or immunosuppressive effects in a JAK2V617F-driven mouse model. [1] The primary end point is reached in 41.9% of patients in the Ruxolitinib group as compared with 0.7% in the placebo group in the double-blind trial of myelofibrosis. Ruxolitinib results in maintaining of reduction in spleen volume and improvement of 50% or more in the total symptom score. [2] A total of 28% of the patients in the Ruxolitinib (15 mg twice daily) group has at least a 35% reduction in spleen volume at week 48 in patients with myelofibrosis, as compared with 0% in the group receiving the best available therapy. The mean palpable spleen length has decreased by 56% with Ruxolitinib but has increased by 4% with the best available therapy at week 48. Patients in the ruxolitinib group has an improvement in overall quality-of-life measures and a reduction in symptoms associated with myelofibrosis. [3]

Protocol

Kinase Assay:[1]
+ Expand

Binding assay:

Recombinant proteins are expressed using Sf21 cells and baculovirus vectors and purified with affinity chromatography. JAK kinase assays use a homogeneous time-resolved fluorescence assay with the peptide substrate (-EQEDEPEGDYFEWLE). Each enzyme reaction is carried out with Ruxolitinib or control, JAK enzyme, 500 nM peptide, adenosine triphosphate (ATP; 1mM), and 2% dimethyl sulfoxide (DMSO) for 1 hour. The 50% inhibitory concentration (IC50) is calculated as INCB018424 concentration required for inhibition of 50% of the fluorescent signal.
Cell Research:[1]
+ Expand
  • Cell lines: Ba/F3 and HEL cells
  • Concentrations: 3 μM
  • Incubation Time: 48 hours
  • Method: Cells are seeded at 2 × 103/well of white bottom 96-well plates, treated with INCB018424 from DMSO stocks (0.2% final DMSO concentration), and incubated for 48 hours at 37 ℃ with 5% CO2. Viability is measured by cellular ATP determination using the Cell-Titer Glo luciferase reagent or viable cell counting. Values are transformed to percent inhibition relative to vehicle control, and IC50 curves are fitted according to nonlinear regression analysis of the data using PRISM GraphPad.
    (Only for Reference)
Animal Research:[1]
+ Expand
  • Animal Models: JAK2V617F-driven mouse model
  • Formulation: 5% dimethyl acetamide, 0.5% methocellulose
  • Dosages: 180 mg/kg
  • Administration: Oral gavage
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 61 mg/mL (199.1 mM)
Water <1 mg/mL
Ethanol <1 mg/mL
In vivo 2% DMSO+30% PEG 300+ddH2O 5mg/mL

* 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.

Chemical Information

Molecular Weight 306.37
Formula

C17H18N6

CAS No. 941678-49-5
Storage powder
in solvent
Synonyms N/A

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Clinical Trial Information

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT01712659 Recruiting T Cell Leukemia, Adult|Leukemia, Adult T-Cell|T Cell Leukemia, HTLV I Associated National Cancer Institute (NCI)|National Institutes of Health Clinical Center (CC) October 3, 2012 Phase 2
NCT02913261 Recruiting Corticosteroid Refractory Acute Graft vs Host Disease Novartis Pharmaceuticals|Novartis February 28, 2017 Phase 3
NCT02973711 Not yet recruiting Leukemia, Chronic Myeloid University of Michigan Cancer Center February 2017 Phase 1|Phase 2
NCT03012230 Not yet recruiting Estrogen Receptor Negative|HER2/Neu Negative|Progesterone Receptor Negative|Stage IV Breast Cancer|Triple-Negative Breast Carcinoma Mayo Clinic|National Cancer Institute (NCI) February 2017 Phase 1
NCT02928978 Not yet recruiting Ductal Carcinoma In Situ|Atypical Lobular Hyperplasia|Atypical Ductal Hyperplasia|Lobular Carcinoma In Situ Julie Nangia|Incyte Corporation|Translational Breast Cancer Research Consortium|Baylor Breast Care Center January 2017 Phase 2
NCT02966353 Not yet recruiting Primary Myelofibrosis (PMF)|Post-Polycythemia Vera-Myelofibrosis (PPV-MF)|Post-Essential Thrombocythemia Myelofibrosis (PET-MF) Novartis Pharmaceuticals|Novartis January 2017 Phase 2

Tech Support

Answers to questions you may have can be found in the inhibitor handling instructions. Topics include how to prepare stock solutions, how to store inhibitors, and issues that need special attention for cell-based assays and animal experiments.

Handling Instructions

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID