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.

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In DMSO USD 208 In stock
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Cited by 36 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

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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 NX;VNXRtU2mwYYPlJGF{e2G7 NWCwVY5zOjBibXnu M{C2[GROW09? MW\Jcohq[mm2aX;uJI9nKEqDS{KgbY4hcHWvYX6gWGYyKGOnbHzzJIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gSXBQNWmwZIXj[YQhW1SDVEWgdIhwe3Cqb4L5cIF1cW:wIIfpeIghUUN3MDDv[kAxNjBzMt88US=> MlXVNlI3QThyOES=
TF1 NWDQV5ROU2mwYYPlJGF{e2G7 NFnoOWgzOCCvaX6= MlPSSG1UVw>? NH[xV|dKdmirYnn0bY9vKG:oIFrBT|EhcW5iaIXtZY4hXEZzIHPlcIx{KGG|c3Xzd4VlKGG|IHnubIljcXSrb36gc4YhUUx4LXnu[JVk\WRiU2TBWFMheGixc4Doc5J6dGG2aX;uJJdqfGhiSVO1NEBw\iByLkCyOO69VQ>? MonxNlI3QThyOES=
Human T cell MX;LbY5ie2ViQYPzZZk> MoPZTY5pcWKrdHnvckBw\iCMQVuzM|EhcW5iaIXtZY4hXCClZXzsd{BmgHC{ZYPzbY5oKEOGMzDhd5Nme3OnZDDhd{BqdmirYnn0bY9vKG:oIFnMNk1{fGmvdXzheIVlKFOWQWS1ZUBxcG:|cHjvdplt[XSrb36ge4l1cCCLQ{WwJI9nKDBwMEKz{txO NIjkdYQzOzV2ME[0PC=>
Human monocyte NHj2eIRMcW6jc3WgRZN{[Xl? M2DW[GlvcGmkaYTpc44hd2ZiSlHLNkBqdiCqdX3hckBud26xY4n0[ZMh\XiycnXzd4lv\yCFREG0JIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gS20uS1OILYP0bY12dGG2ZXSgV3RCXDWjIIDoc5NxcG:{eXzheIlwdiC5aYToJGlEPTBib3[gNE4xOjcQvF2= NFfDZZYzOzV2ME[0PC=>
Human monocyte M171UmtqdmG|ZTDBd5NigQ>? Mn3WTY5pcWKrdHnvckBw\iCMQVuyM|EhcW5iaIXtZY4hdW:wb3P5eIV{KGW6cILld5NqdmdiQ1SxOEBie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGlHVmejbX3hMZN1cW23bHH0[YQhW1SDVEGgdIhwe3Cqb4L5cIF1cW:wIIfpeIghUUN3MDDv[kAxNjB|Md88US=> MV[yN|U1ODZ2OB?=
HEL NIHaTGVEgXSxdH;4bYMhSXO|YYm= MWG1JO69VQ>? MU[0PEBp NHfhV49EgXSxdH;4bYMhcW6mZYi9NVIvOiV? Mm\kNlU6OzF|NEm=
SET-2 MofKR5l1d3SxeHnjJGF{e2G7 NV\OeplnPSEQvF2= MlHUOFghcA>? NGnqZXpEgXSxdH;4bYMhcW6mZYi9NVgvPyV? MV6yOVk{OTN2OR?=
HT93A NHTlbWxIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MVmzNlAhdk1? MXW1JIQ> MkHpSG1UVw>? MmD2TY5pcWKrdHnvckBw\iCJQ2OtSkBqdmS3Y3XkJIdz[W63bH;jfZRq[yCmaX\m[ZJmdnSrYYTpc44> NWT3RmF5OjV6MEW5OlI>
CMK NVzlfoFoT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NWnJO5l2UW6qaXLpeIlwdiCxZjDDUWsh[2G{conpcochfGinIFrBT|NCPTd{VjDteZRifGmxbjDj[YxtKHC{b3zp[oVz[XSrb36= MWSyOVM2OjF{NB?=
CMK NEHORm9Iem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NFLxW|ZKdmirYnn0bY9vKG:oIFPNT{Bk[XK{eXnu[{B1cGViSlHLN2E3O0RibYX0ZZRqd25iY3XscEBxem:uaX\ldoF1cW:wIIfpeIghUUN3MDDv[kAxNjF4MzFOwG0> Ml3TNlU{PTJzMkS=
CMK MmHiS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NYSzflBkUW6qaXLpeIlwdiCxZjDDUWsh[2G{conpcochfGinIGfUJGpCUyClZXzsJJBzd2yrZnXyZZRqd25id3n0bEBKSzVyIH;mJFAvODd3IN88US=> Mn6yNlU{PTJzMkS=
NCI-H460 MXPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NVPrc21XTE2VTx?= MmrUTWM2OD1yLkGzJO69VQ>? MlflNlUzOTN4N{C=
NCI-H358 NVT5NIt[T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MmHVSG1UVw>? NHvGdZNKSzVyPUCuNUDPxE1? MmHGNlUzOTN4N{C=
A549 Mm\CS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M{D6ZmROW09? Mn\YTWM2OD1yLkC0JO69VQ>? NV\xN2djOjV{MUO2O|A>
A549/DDP M2fzTWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NYrPb25NTE2VTx?= M13sVGlEPTB;MD6yNkDPxE1? MU[yOVIyOzZ5MB?=
NCI-H1299 MWXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M3TnNGROW09? MkDoTWM2OD1yLkK4JO69VQ>? MUmyOVIyOzZ5MB?=
NCI-H2347 MoGzS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NX7uT4s2TE2VTx?= M2\BW2lEPTB;MD6xO{DPxE1? MlHWNlUzOTN4N{C=
A549/DDP MVzGeY5kfGmxbjDBd5NigQ>? NWr5cZA4OzBibl2= NHfPOIk1QCCq M{n3eGROW09? MV7Ec5dvNXKnZ4XsZZRqd25ib3[gV3RCXDNicHjvd5Bpd3K7bHH0bY9v M4jnc|I2OjF|Nkew
NCI-H1299 M16xNWZ2dmO2aX;uJGF{e2G7 M{DaTVMxKG6P MmPEOFghcA>? NVHJZ3RUTE2VTx?= MljkSI94di2{ZXf1cIF1cW:wIH;mJHNVSVR|IIDoc5NxcG:{eXzheIlwdg>? NV\ENZoxOjV{MUO2O|A>
NCI-H2347 M3rkUmZ2dmO2aX;uJGF{e2G7 MlrPN|Ahdk1? MV20PEBp NFfITnRFVVOR MljSSIVkemWjc3WgbY4hSmOuMjDlfJBz\XO|aX;u NIDVdGYzPTJzM{[3NC=>
A549/DDP MlPQRZBweHSxc3nzJGF{e2G7 Mn21N|Ahdk1? MXW0PEBp M3jFWmROW09? MoP0TY5lfWO2aX;uJI9nKGGyb4D0c5Nqew>? MW[yOVIyOzZ5MB?=
NCI-H1299 NHW2dFFCeG:ydH;zbZMhSXO|YYm= M2nkPVMxKG6P MXS0PEBp M1\PbGROW09? Ml3STY5lfWO2aX;uJI9nKGGyb4D0c5Nqew>? Mn;wNlUzOTN4N{C=
NCI-H2347 MnHtRZBweHSxc3nzJGF{e2G7 Mn\EN|Ahdk1? NVztO2tqPDhiaB?= NF\uOo9FVVOR MXjJcoR2[3Srb36gc4Yh[XCxcITvd4l{ Ml\aNlUzOTN4N{C=
Hep3B M3vU[GZ2dmO2aX;uJGF{e2G7 M{XN[FEh|ryP MXuxOkBp M3PJR2ROW09? MnPhTY1x[Wm{ZYOgeIhmKGOjcHHjbZR6KG:oIFnIR2Eu[XO|b3PpZZRm\CCpcEGzNEBufXSjboTzJJRwKGGldHn2[UBUXEGWMzD3bZRpKEmFNUCgc4YhhjVyIN88US=> NIG3VnAzPDVyMU[4PS=>
HepG2 NI\rTXFHfW6ldHnvckBCe3OjeR?= MoDRNUDPxE1? M3LyN|E3KGh? MUnEUXNQ NGDrdZFKdXCjaYLld{B1cGViY3HwZYNqfHlib3[gTWhESS2jc4PvZ4lifGWmIHfwNVMxKG23dHHueJMhfG9ic3nncoFtKHSxIGPURXQ{ M{KxWVI1PTBzNki5
Huh7 M4jpOGZ2dmO2aX;uJGF{e2G7 MnnkNUDPxE1? MlHnNVYhcA>? MY\EUXNQ MmnVTY1x[Wm{ZYOgeIhmKGOjcHHjbZR6KG:oIFnIR2Eu[XO|b3PpZZRm\CCpcEGzNEBufXSjboTzJJRwKHOrZ37hcEB1dyCVVFHUNy=> NGq4NXYzPDVyMU[4PS=>
BaF3 MlzlT4lv[XOnIFHzd4F6 M2LLWlgxKG6P M13ndlYhcA>? M3XRZmROW09? NXvDUZlnWmWmdXPld{B1cGVicHjvd5Bpd3K7bHH0bY9vKG:owrDTWGFVPSCrbjDKRWszXjZzN1[tcZV1[XSnZDDCRWY{NUWST2KgZ4VtdA>? NWDqT|NsOjR{M{e3PVE>
DLD-1 Mnr4T4lv[XOnIFHzd4F6 NICwT4QzPSEQvF2= M4jMXFQ5KGh? NYTJdppCTE2VTx?= Mon0TY5pcWKrdHnvckBw\iCMQVuxJJBpd3OyaH;yfYxifGmxbh?= MWKyOFA2ODV3MB?=
RKO MXjLbY5ie2ViQYPzZZk> MmqwNlUh|ryP M4jlUFQ5KGh? MoLjSG1UVw>? MYTJcohq[mm2aX;uJI9nKEqDS{GgdIhwe3Cqb4L5cIF1cW:w MYWyOFA2ODV3MB?=
DLD-1 NWXU[nZmU2mwYYPlJGF{e2G7 MlnWNlUh|ryP M{HD[|Q5KGh? NFX0U|hFVVOR MmrJTY5pcWKrdHnvckBw\iCMQVuyJJBpd3OyaH;yfYxifGmxbh?= MUGyOFA2ODV3MB?=
RKO NGnTPG5McW6jc3WgRZN{[Xl? NUPHNGZlOjVizszN NVThc|h2PDhiaB?= Ml\ZSG1UVw>? M2T2foRw\XNibn;0JIlvcGmkaYSgTmFMOSCyaH;zdIhwenmuYYTpc44> MofzNlQxPTB3NUC=
DLD-1 M{TOeWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 M1LGV|UxKM7:TR?= MmS3OFghcA>? MWHEUXNQ MYfJR|UxRTF3LkWxJO69VQ>? M3XKNVI1ODVyNUWw
RKO NH3p[HhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NH:0fmQ2OCEQvF2= MmTROFghcA>? NW\KV3Z1TE2VTx?= MX\JR|UxRTF2Lke2JO69VQ>? MUKyOFA2ODV3MB?=
DLD-1 NHvzUpFCeG:ydH;zbZMhSXO|YYm= MYqyOUDPxE1? NGTzfpY1QCCq MVvEUXNQ NX7VSnFMUW6mdXPld{BieG:ydH;zbZMh[nliYXP0bZZifGmwZzDjZZNx[XOnIEO= M3HPclI1ODVyNUWw
RKO M{nPVWFxd3C2b4Ppd{BCe3OjeR?= NHzLSXAzPSEQvF2= Mn;nOFghcA>? M2DMXWROW09? MU\JcoR2[2W|IHHwc5B1d3OrczDifUBi[3SrdnH0bY5oKGOjc4Dhd4UhOw>? MlT2NlQxPTB3NUC=
HuH7 Ml;DS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MYm1NEDPxE1? MY[0PEBp M3vqcmROW09? NHj1cok,QDJnIILl[JVkfGmxbh?= M1\od|I{QTRzOEOy
SNU182 NXLSbXU3T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MUi1NEDPxE1? MoDCOFghcA>? NGLjR41FVVOR MljEQlY1LSC{ZXT1Z5Rqd25? MWqyN|k1OTh|Mh?=
SNU423 M2X2UGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MVm1NEDPxE1? MXi0PEBp M{PoWGROW09? MXW+PFEmKHKnZIXjeIlwdg>? M4jUNVI{QTRzOEOy
HuH7 NVTaTWE5TnWwY4Tpc44hSXO|YYm= M1[0SVUxKM7:TR?= M2TEUFI1KGh? NHryeVJFVVOR Ml;PTY5pcWKrdHnvckBw\iCVVFHUNUBidmRiU2TBWFMheGixc4Doc5J6dGG2aX;uJJNq\26rZnnjZY51dHl? NVy0O|RtOjN7NEG4N|I>
SNU182 MVrGeY5kfGmxbjDBd5NigQ>? NWHlcnhvPTBizszN MX2yOEBp MY\EUXNQ NYDuO4NiUW6qaXLpeIlwdiCxZjDTWGFVOSCjbnSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKHOrZ37p[olk[W62bIm= M1zhZ|I{QTRzOEOy
SNU423 MWjGeY5kfGmxbjDBd5NigQ>? MV21NEDPxE1? MoHtNlQhcA>? NXPFZoN3TE2VTx?= Mnz5TY5pcWKrdHnvckBw\iCVVFHUNUBidmRiU2TBWFMheGixc4Doc5J6dGG2aX;uJJNq\26rZnnjZY51dHl? MWOyN|k1OTh|Mh?=

... 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

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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.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID