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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Cited by 36 Publications

5 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.

    INCB018424 administration reverses the protective effects of ALA on ONC retinas. A-C: Representative micrographs (200× magnification) of retinal whole mounts obtained from three groups stained with anti-RNA-binding protein with multiple splicing (Rbpms) antibody (green). Sampling location: 2 mm temporal to the optic disc. Sampling field size: 439 × 330 μm2 (20× objective lens). Scale bar: 50 μm. D: Quantitative analysis of Rbpms-positive cells under different experimental conditions (mean ± standard error of the mean [SEM], n = 6 per group). The average number of Rbpms-positive cells/mm2 was calculated. ONC = optic nerve crush animal; ALA-ONC = alpha lipoic acid (ALA) animal pretreated 1 day before ONC; ALA-ONC+I = ALA animal pretreated 1 day before ONC, followed by INCB018424. *** p<0.001, ** p<0.01 compared to the ONC group, ### p<0.001 compared to the ALA-ONC group at the same time point

    Mol Vis, 2016, 22:1122-1136. 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.

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Choose Selective JAK Inhibitors

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 NGTiPIRMcW6jc3WgRZN{[Xl? M1LOOlIxKG2rbh?= NHvWXJdFVVOR NGDIR25KdmirYnn0bY9vKG:oIFrBT|IhcW5iaIXtZY4hXEZzIHPlcIx{KGG|c3Xzd4VlKGG|IHnubIljcXSrb36gc4YhTVCRLXnu[JVk\WRiU2TBWFUheGixc4Doc5J6dGG2aX;uJJdqfGhiSVO1NEBw\iByLkCxNu69VQ>? M1TnfFIzPjl6MEi0
TF1 MUDLbY5ie2ViQYPzZZk> MoLMNlAhdWmw M3jIPGROW09? MmHDTY5pcWKrdHnvckBw\iCMQVuxJIlvKGi3bXHuJHRHOSClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGlNPi2rbnT1Z4VlKFOWQWSzJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMkVOwG0> NHHVSY4zOjZ7OEC4OC=>
Human T cell NF65VYdMcW6jc3WgRZN{[Xl? NVLyRXZoUW6qaXLpeIlwdiCxZjDKRWs{NzFiaX6gbJVu[W5iVDDj[YxteyCneIDy[ZN{cW6pIFPEN{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGlNOi2|dHnteYxifGWmIGPURXQ2[SCyaH;zdIhwenmuYYTpc44hf2m2aDDJR|UxKG:oIECuNFI{|ryP NXf2Z204OjN3NEC2OFg>
Human monocyte MVnLbY5ie2ViQYPzZZk> M3m5fWlvcGmkaYTpc44hd2ZiSlHLNkBqdiCqdX3hckBud26xY4n0[ZMh\XiycnXzd4lv\yCFREG0JIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gS20uS1OILYP0bY12dGG2ZXSgV3RCXDWjIIDoc5NxcG:{eXzheIlwdiC5aYToJGlEPTBib3[gNE4xOjcQvF2= MnrSNlM2PDB4NEi=
Human monocyte M1LTcWtqdmG|ZTDBd5NigQ>? M3HhdGlvcGmkaYTpc44hd2ZiSlHLNk8yKGmwIHj1cYFvKG2xbn;jfZRmeyCneIDy[ZN{cW6pIFPENVQh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBKTk6pYX3tZU1{fGmvdXzheIVlKFOWQWSxJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yM{JOwG0> MYOyN|U1ODZ2OB?=
HEL M1vkXmN6fG:2b4jpZ{BCe3OjeR?= M2raXFUh|ryP MUC0PEBp M4C0bWN6fG:2b4jpZ{BqdmSneE2xNk4zLQ>? NHLLT|QzPTl|MUO0PS=>
SET-2 NUjyRYtCS3m2b4TvfIlkKEG|c3H5 MX[1JO69VQ>? NFTTSGQ1QCCq MkW3R5l1d3SxeHnjJIlv\GW6PUG4Mlcm MnmwNlU6OzF|NEm=
HT93A Mm\uS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M4TQUlMzOCCwTR?= M4iyeFUh\A>? NX\aOnNjTE2VTx?= NVPFTFlTUW6qaXLpeIlwdiCxZjDHR3MuTiCrbnT1Z4VlKGe{YX71cI9kgXSrYzDkbYZn\XKnboTpZZRqd25? MX6yOVgxPTl4Mh?=
CMK M3fLV2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NGWwR|dKdmirYnn0bY9vKG:oIFPNT{Bk[XK{eXnu[{B1cGViSlHLN2E2PzKYIH31eIF1cW:wIHPlcIwheHKxbHnm[ZJifGmxbh?= MXmyOVM2OjF{NB?=
CMK M3rReWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NFHUOJBKdmirYnn0bY9vKG:oIFPNT{Bk[XK{eXnu[{B1cGViSlHLN2E3O0RibYX0ZZRqd25iY3XscEBxem:uaX\ldoF1cW:wIIfpeIghUUN3MDDv[kAxNjF4MzFOwG0> NWPtc4VGOjV|NUKxNlQ>
CMK MV\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NX;QSFdIUW6qaXLpeIlwdiCxZjDDUWsh[2G{conpcochfGinIGfUJGpCUyClZXzsJJBzd2yrZnXyZZRqd25id3n0bEBKSzVyIH;mJFAvODd3IN88US=> NFrUOmUzPTN3MkGyOC=>
NCI-H460 NYP3WIhuT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MYDEUXNQ NFO0WFdKSzVyPUCuNVMh|ryP MWCyOVIyOzZ5MB?=
NCI-H358 MXLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NY\ZXY5nTE2VTx?= NFTkTHpKSzVyPUCuNUDPxE1? MlPDNlUzOTN4N{C=
A549 NFnjbnhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NWXwVHJ2TE2VTx?= NF7ZemZKSzVyPUCuNFQh|ryP M2r2b|I2OjF|Nkew
A549/DDP M4L6ZWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NIXidJVFVVOR MWTJR|UxRTBwMkKg{txO M3n6[|I2OjF|Nkew
NCI-H1299 NVro[Hd6T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MkHDSG1UVw>? M1X6bGlEPTB;MD6yPEDPxE1? M4nVVFI2OjF|Nkew
NCI-H2347 M3i2cWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NUT5UpFvTE2VTx?= MkXUTWM2OD1yLkG3JO69VQ>? MmnHNlUzOTN4N{C=
A549/DDP NXTZS49sTnWwY4Tpc44hSXO|YYm= MUizNEBvVQ>? M4n0VlQ5KGh? NYDyOZJ4TE2VTx?= MWjEc5dvNXKnZ4XsZZRqd25ib3[gV3RCXDNicHjvd5Bpd3K7bHH0bY9v MXiyOVIyOzZ5MB?=
NCI-H1299 NHvhOolHfW6ldHnvckBCe3OjeR?= NYf4fIJTOzBibl2= NI[0WW81QCCq NHTIVW1FVVOR M4XaZWRwf25vcnXneYxifGmxbjDv[kBUXEGWMzDwbI9{eGixconsZZRqd25? MlfhNlUzOTN4N{C=
NCI-H2347 NIDrSW1HfW6ldHnvckBCe3OjeR?= MlLMN|Ahdk1? M3mxRlQ5KGh? MkPFSG1UVw>? Mn[wSIVkemWjc3WgbY4hSmOuMjDlfJBz\XO|aX;u NHfu[lIzPTJzM{[3NC=>
A549/DDP NVm5S21XSXCxcITvd4l{KEG|c3H5 NIjwR5Q{OCCwTR?= NXnPWVNRPDhiaB?= NF3YVWZFVVOR MmjLTY5lfWO2aX;uJI9nKGGyb4D0c5Nqew>? MVuyOVIyOzZ5MB?=
NCI-H1299 NIPjSFNCeG:ydH;zbZMhSXO|YYm= M3\0S|MxKG6P NGLx[pA1QCCq M2CzVGROW09? NFjhXoZKdmS3Y4Tpc44hd2ZiYYDvdJRwe2m| M1;0eFI2OjF|Nkew
NCI-H2347 M1OyVWFxd3C2b4Ppd{BCe3OjeR?= NVnGe4hHOzBibl2= MlvsOFghcA>? NUDMRoRoTE2VTx?= NYO3fFN5UW6mdXP0bY9vKG:oIHHwc5B1d3Orcx?= NYq1TFdxOjV{MUO2O|A>
Hep3B NXfJbGF2TnWwY4Tpc44hSXO|YYm= NVHvNm5QOSEQvF2= MnXXNVYhcA>? Mn3JSG1UVw>? NXPoVGc{UW2yYXny[ZMhfGinIHPhdIFkcXS7IH;mJGlJS0FvYYPzc4Nq[XSnZDDndFE{OCCvdYThcpR{KHSxIHHjeIl3\SCVVFHUN{B4cXSqIFnDOVAhd2ZifkWwJO69VQ>? MkjQNlQ2ODF4OEm=
HepG2 Ml\oSpVv[3Srb36gRZN{[Xl? M3nZU|Eh|ryP MXuxOkBp MV\EUXNQ MVXJcZBicXKnczD0bIUh[2GyYXPpeJkhd2ZiSVjDRU1ie3OxY3nheIVlKGeyMUOwJI12fGGwdIOgeI8he2mpbnHsJJRwKFOWQWSz MmnPNlQ2ODF4OEm=
Huh7 MWjGeY5kfGmxbjDBd5NigQ>? MmmyNUDPxE1? MWmxOkBp MnTjSG1UVw>? NUju[|dlUW2yYXny[ZMhfGinIHPhdIFkcXS7IH;mJGlJS0FvYYPzc4Nq[XSnZDDndFE{OCCvdYThcpR{KHSxIIPp[45idCC2bzDTWGFVOw>? MYOyOFUxOTZ6OR?=
BaF3 NHvyNlZMcW6jc3WgRZN{[Xl? NWPhXlRtQDBibl2= MkTtOkBp MUTEUXNQ NEnvU2xT\WS3Y3XzJJRp\SCyaH;zdIhwenmuYYTpc44hd2cEoGPURXQ2KGmwIFrBT|JXPjF5Rj3teZRifGWmIFLBSlMuTVCRUjDj[Yxt MYGyOFI{Pzd7MR?=
DLD-1 NWTWZ4t3U2mwYYPlJGF{e2G7 M1zZe|I2KM7:TR?= MVG0PEBp NYXDZ|IxTE2VTx?= NEDCUZVKdmirYnn0bY9vKG:oIFrBT|EheGixc4Doc5J6dGG2aX;u MlTvNlQxPTB3NUC=
RKO NFjNPXlMcW6jc3WgRZN{[Xl? NGHTOnYzPSEQvF2= MWm0PEBp M1zRTWROW09? M2e4[WlvcGmkaYTpc44hd2ZiSlHLNUBxcG:|cHjvdplt[XSrb36= NIPhVHYzPDB3MEW1NC=>
DLD-1 NHe2[ldMcW6jc3WgRZN{[Xl? M{nLd|I2KM7:TR?= NVP6PXNzPDhiaB?= NGS0WmlFVVOR NWLuSmdqUW6qaXLpeIlwdiCxZjDKRWszKHCqb4PwbI9zgWyjdHnvci=> MW[yOFA2ODV3MB?=
RKO NUXrN4s5U2mwYYPlJGF{e2G7 MVyyOUDPxE1? NX;L[YIyPDhiaB?= Ml60SG1UVw>? NXHEWYR{\G:nczDuc5QhcW6qaXLpeEBLSUtzIIDoc5NxcG:{eXzheIlwdg>? MlnJNlQxPTB3NUC=
DLD-1 NGftcIhIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M1K0NFUxKM7:TR?= MlTYOFghcA>? MUnEUXNQ NG\TZndKSzVyPUG1MlUyKM7:TR?= MV:yOFA2ODV3MB?=
RKO NEPWdGdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MXG1NEDPxE1? NVTtNWRWPDhiaB?= MWrEUXNQ MXHJR|UxRTF2Lke2JO69VQ>? MVuyOFA2ODV3MB?=
DLD-1 M4PyUmFxd3C2b4Ppd{BCe3OjeR?= MlPnNlUh|ryP NF[1SGY1QCCq M4DF[WROW09? M2npbmlv\HWlZYOgZZBweHSxc3nzJIJ6KGGldHn2ZZRqdmdiY3HzdIF{\SB| MY[yOFA2ODV3MB?=
RKO NF3DVZpCeG:ydH;zbZMhSXO|YYm= NHTPRoYzPSEQvF2= NHLsTW41QCCq M{m3bmROW09? NGroc|NKdmS3Y3XzJIFxd3C2b4Ppd{BjgSCjY4TpeoF1cW6pIHPhd5Bie2ViMx?= MY[yOFA2ODV3MB?=
HuH7 NYrsNlk6T3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MnPVOVAh|ryP NY\oS4RmPDhiaB?= MXrEUXNQ NXHtXGF3Rjh{JTDy[YR2[3Srb36= MVmyN|k1OTh|Mh?=
SNU182 MXnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NFrFfpA2OCEQvF2= MlXUOFghcA>? M2PncGROW09? M3XQO|43PCVicnXkeYN1cW:w M1rXblI{QTRzOEOy
SNU423 Mm[3S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M4H6T|UxKM7:TR?= NXH2Wnc{PDhiaB?= NF3xXVJFVVOR NX3Rd3p7RjhzJTDy[YR2[3Srb36= Mn;uNlM6PDF6M{K=
HuH7 MkTrSpVv[3Srb36gRZN{[Xl? M{DxT|UxKM7:TR?= NUm4VWg6OjRiaB?= NYnQOIFrTE2VTx?= MV;Jcohq[mm2aX;uJI9nKFOWQWSxJIFv\CCVVFHUN{BxcG:|cHjvdplt[XSrb36gd4lodmmoaXPhcpRtgQ>? M{PwV|I{QTRzOEOy
SNU182 NXrPdZVCTnWwY4Tpc44hSXO|YYm= NUDjfYFLPTBizszN Mm[5NlQhcA>? NIj4fpNFVVOR NYrZNodnUW6qaXLpeIlwdiCxZjDTWGFVOSCjbnSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKHOrZ37p[olk[W62bIm= Mo[zNlM6PDF6M{K=
SNU423 M4jO[GZ2dmO2aX;uJGF{e2G7 MYO1NEDPxE1? NWHUUYNlOjRiaB?= MljPSG1UVw>? NWLyVW5pUW6qaXLpeIlwdiCxZjDTWGFVOSCjbnSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKHOrZ37p[olk[W62bIm= MmWwNlM6PDF6M{K=

... 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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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID