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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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 MXPLbY5ie2ViQYPzZZk> NXvXbHRROjBibXnu MWHEUXNQ MnTJTY5pcWKrdHnvckBw\iCMQVuyJIlvKGi3bXHuJHRHOSClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGVRVy2rbnT1Z4VlKFOWQWS1JJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMUNOwG0> MXyyNlY6QDB6NB?=
TF1 M4TURmtqdmG|ZTDBd5NigQ>? NHnxW48zOCCvaX6= MnzhSG1UVw>? M1\kPGlvcGmkaYTpc44hd2ZiSlHLNUBqdiCqdX3hckBVTjFiY3XscJMh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBKVDZvaX7keYNm\CCVVFHUN{BxcG:|cHjvdplt[XSrb36ge4l1cCCLQ{WwJI9nKDBwMEK0{txO MnG4NlI3QThyOES=
Human T cell MWjLbY5ie2ViQYPzZZk> M{XhTGlvcGmkaYTpc44hd2ZiSlHLN{8yKGmwIHj1cYFvKFRiY3XscJMh\XiycnXzd4lv\yCFREOgZZN{\XO|ZXSgZZMhcW6qaXLpeIlwdiCxZjDJUFIue3SrbYXsZZRm\CCVVFHUOYEheGixc4Doc5J6dGG2aX;uJJdqfGhiSVO1NEBw\iByLkCyN:69VQ>? MU[yN|U1ODZ2OB?=
Human monocyte NIPhUJVMcW6jc3WgRZN{[Xl? NXS2R3UxUW6qaXLpeIlwdiCxZjDKRWszKGmwIHj1cYFvKG2xbn;jfZRmeyCneIDy[ZN{cW6pIFPENVQh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBIVS2FU1[td5RqdXWuYYTl[EBUXEGWNXGgdIhwe3Cqb4L5cIF1cW:wIIfpeIghUUN3MDDv[kAxNjB{Nt88US=> MoXKNlM2PDB4NEi=
Human monocyte MVHLbY5ie2ViQYPzZZk> NGnTdWZKdmirYnn0bY9vKG:oIFrBT|IwOSCrbjDoeY1idiCvb37vZ5l1\XNiZYjwdoV{e2mwZzDDSFE1KGG|c3Xzd4VlKGG|IHnubIljcXSrb36gc4YhUU[QZ3HtcYEue3SrbYXsZZRm\CCVVFHUNUBxcG:|cHjvdplt[XSrb36ge4l1cCCLQ{WwJI9nKDBwMEOx{txO NUn5b|J6OjN3NEC2OFg>
HEL MWTDfZRwfG:6aXOgRZN{[Xl? NIXpSog2KM7:TR?= M4\2SlQ5KGh? NWLQem5CS3m2b4TvfIlkKGmwZHX4QVEzNjJn M1jMTlI2QTNzM{S5
SET-2 Ml3SR5l1d3SxeHnjJGF{e2G7 MX61JO69VQ>? NIjmb|E1QCCq NVHwV3dXS3m2b4TvfIlkKGmwZHX4QVE5Njdn M3X2[lI2QTNzM{S5
HT93A MU\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? M{\ES|MzOCCwTR?= M1LkOlUh\A>? MVLEUXNQ MXzJcohq[mm2aX;uJI9nKEeFUz3GJIlv\HWlZXSg[5JidnWub3P5eIlkKGSrZn\ldoVvfGmjdHnvci=> MUGyOVgxPTl4Mh?=
CMK MkDPS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M33PXmlvcGmkaYTpc44hd2ZiQ13LJINienK7aX7nJJRp\SCMQVuzRVU4OlZibYX0ZZRqd25iY3XscEBxem:uaX\ldoF1cW:w M37qZVI2OzV{MUK0
CMK NGLEUIZIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MljUTY5pcWKrdHnvckBw\iCFTVugZ4FzenmrbnegeIhmKEqDS{PBOlNFKG23dHH0bY9vKGOnbHygdJJwdGmoZYLheIlwdiC5aYToJGlEPTBib3[gNE4yPjNizszN MYeyOVM2OjF{NB?=
CMK MWXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NVLneJhmUW6qaXLpeIlwdiCxZjDDUWsh[2G{conpcochfGinIGfUJGpCUyClZXzsJJBzd2yrZnXyZZRqd25id3n0bEBKSzVyIH;mJFAvODd3IN88US=> NW\NboxkOjV|NUKxNlQ>
NCI-H460 NFyze2VIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M3PEUmROW09? MYrJR|UxRTBwMUOg{txO MWmyOVIyOzZ5MB?=
NCI-H358 NHvTSnlIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MV7EUXNQ NIq3eYJKSzVyPUCuNUDPxE1? NVHrRnBmOjV{MUO2O|A>
A549 NIrjOlRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NVLnc242TE2VTx?= MYPJR|UxRTBwMESg{txO NUHXRVBvOjV{MUO2O|A>
A549/DDP NUHHZoxpT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MmrySG1UVw>? MlTUTWM2OD1yLkKyJO69VQ>? MnnGNlUzOTN4N{C=
NCI-H1299 NXziOI9uT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= MV\EUXNQ MmP2TWM2OD1yLkK4JO69VQ>? NULPZmZrOjV{MUO2O|A>
NCI-H2347 M{fJSWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHTWWY9FVVOR M{HYXWlEPTB;MD6xO{DPxE1? NEHqN2UzPTJzM{[3NC=>
A549/DDP M2LX[WZ2dmO2aX;uJGF{e2G7 NXXXcI0yOzBibl2= Ml;hOFghcA>? MYnEUXNQ NELicoVFd3ewLYLl[5Vt[XSrb36gc4YhW1SDVEOgdIhwe3Cqb4L5cIF1cW:w MXWyOVIyOzZ5MB?=
NCI-H1299 NF;pN5FHfW6ldHnvckBCe3OjeR?= M1H5flMxKG6P NIfmUXQ1QCCq NU\BeYN1TE2VTx?= M1rQ[WRwf25vcnXneYxifGmxbjDv[kBUXEGWMzDwbI9{eGixconsZZRqd25? MlK2NlUzOTN4N{C=
NCI-H2347 Mkm5SpVv[3Srb36gRZN{[Xl? MUmzNEBvVQ>? M2nLNlQ5KGh? M1TidWROW09? NYH2PGxwTGWlcnXhd4UhcW5iQnPsNkBmgHC{ZYPzbY9v MlPLNlUzOTN4N{C=
A549/DDP NV\aOpFVSXCxcITvd4l{KEG|c3H5 NIPvelE{OCCwTR?= NIjNfVA1QCCq MVzEUXNQ M4rIU2lv\HWldHnvckBw\iCjcH;weI9{cXN? NH[zOogzPTJzM{[3NC=>
NCI-H1299 NXvZem1iSXCxcITvd4l{KEG|c3H5 MmG0N|Ahdk1? MnXvOFghcA>? MVnEUXNQ NYfqVHB7UW6mdXP0bY9vKG:oIHHwc5B1d3Orcx?= MnzmNlUzOTN4N{C=
NCI-H2347 NWraS3JzSXCxcITvd4l{KEG|c3H5 NFqxbVk{OCCwTR?= M4i5e|Q5KGh? MUDEUXNQ MoT0TY5lfWO2aX;uJI9nKGGyb4D0c5Nqew>? MkDXNlUzOTN4N{C=
Hep3B M{HhcGZ2dmO2aX;uJGF{e2G7 NHnNclkyKM7:TR?= NFTEc2kyPiCq MnjvSG1UVw>? MVzJcZBicXKnczD0bIUh[2GyYXPpeJkhd2ZiSVjDRU1ie3OxY3nheIVlKGeyMUOwJI12fGGwdIOgeI8h[WO2aY\lJHNVSVR|IIfpeIghUUN3MDDv[kB,PTBizszN MXqyOFUxOTZ6OR?=
HepG2 MnvjSpVv[3Srb36gRZN{[Xl? NV3ibIc1OSEQvF2= MmfLNVYhcA>? M4\mRWROW09? MnXRTY1x[Wm{ZYOgeIhmKGOjcHHjbZR6KG:oIFnIR2Eu[XO|b3PpZZRm\CCpcEGzNEBufXSjboTzJJRwKHOrZ37hcEB1dyCVVFHUNy=> NX3JV5BDOjR3MEG2PFk>
Huh7 NGXabohHfW6ldHnvckBCe3OjeR?= NXznSZQ5OSEQvF2= M{njZVE3KGh? MnfKSG1UVw>? NYj4Z5JQUW2yYXny[ZMhfGinIHPhdIFkcXS7IH;mJGlJS0FvYYPzc4Nq[XSnZDDndFE{OCCvdYThcpR{KHSxIIPp[45idCC2bzDTWGFVOw>? MXuyOFUxOTZ6OR?=
BaF3 Mn\UT4lv[XOnIFHzd4F6 M3L6O|gxKG6P MX:2JIg> M2PmO2ROW09? M13uPXJm\HWlZYOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\sLiU2TBWFUhcW5iSlHLNnY3OTeILX31eIF1\WRiQlHGN{1GWE:UIHPlcIw> M{W0VlI1OjN5N{mx
DLD-1 MWDLbY5ie2ViQYPzZZk> NVXZUJB2OjVizszN M3zzVFQ5KGh? NUnKZY1kTE2VTx?= MmHZTY5pcWKrdHnvckBw\iCMQVuxJJBpd3OyaH;yfYxifGmxbh?= MVGyOFA2ODV3MB?=
RKO MVnLbY5ie2ViQYPzZZk> MUCyOUDPxE1? MX:0PEBp NX7qZ3p1TE2VTx?= M4mwOGlvcGmkaYTpc44hd2ZiSlHLNUBxcG:|cHjvdplt[XSrb36= NXTLN3hVOjRyNUC1OVA>
DLD-1 NIe2U49McW6jc3WgRZN{[Xl? NEKyeokzPSEQvF2= Mlf6OFghcA>? MYTEUXNQ Mmf5TY5pcWKrdHnvckBw\iCMQVuyJJBpd3OyaH;yfYxifGmxbh?= MlfNNlQxPTB3NUC=
RKO M3zFcmtqdmG|ZTDBd5NigQ>? NFvWVI0zPSEQvF2= NULoNYo1PDhiaB?= MVHEUXNQ M4DjUYRw\XNibn;0JIlvcGmkaYSgTmFMOSCyaH;zdIhwenmuYYTpc44> MlvJNlQxPTB3NUC=
DLD-1 Mn3mS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M1nscFUxKM7:TR?= Mk\tOFghcA>? MlfQSG1UVw>? NUnCNIRDUUN3ME2xOU42OSEQvF2= MWSyOFA2ODV3MB?=
RKO MY\Hdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NUX0bJliPTBizszN M{DVXFQ5KGh? NXfscJRHTE2VTx?= NIfDNVFKSzVyPUG0Mlc3KM7:TR?= M{nhTlI1ODVyNUWw
DLD-1 M1;sc2Fxd3C2b4Ppd{BCe3OjeR?= NILpWnEzPSEQvF2= MY[0PEBp NVLi[XBSTE2VTx?= MYHJcoR2[2W|IHHwc5B1d3OrczDifUBi[3SrdnH0bY5oKGOjc4Dhd4UhOw>? M2XOeVI1ODVyNUWw
RKO NEDrcG9CeG:ydH;zbZMhSXO|YYm= NXLXfWVIOjVizszN MXy0PEBp NULScnl5TE2VTx?= MUjJcoR2[2W|IHHwc5B1d3OrczDifUBi[3SrdnH0bY5oKGOjc4Dhd4UhOw>? M1v3PFI1ODVyNUWw
HuH7 MYPHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MWS1NEDPxE1? NWHpbmJ[PDhiaB?= NIL2c5hFVVOR M1jvRV45OiVicnXkeYN1cW:w Mo\zNlM6PDF6M{K=
SNU182 MnLuS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M{XjTlUxKM7:TR?= NUXB[|U2PDhiaB?= M4LLPWROW09? M2Dsel43PCVicnXkeYN1cW:w NWG2OlVsOjN7NEG4N|I>
SNU423 M2fUc2dzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHzpRYU2OCEQvF2= NV[1WZN{PDhiaB?= MVnEUXNQ MX[+PFEmKHKnZIXjeIlwdg>? MYOyN|k1OTh|Mh?=
HuH7 NHzVWZRHfW6ldHnvckBCe3OjeR?= M4m1R|UxKM7:TR?= MkXDNlQhcA>? NH3Oe|JFVVOR NWjNTIFNUW6qaXLpeIlwdiCxZjDTWGFVOSCjbnSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKHOrZ37p[olk[W62bIm= MlGxNlM6PDF6M{K=
SNU182 M3X5WGZ2dmO2aX;uJGF{e2G7 M1LEZVUxKM7:TR?= NHPmfYUzPCCq NVrxZW1xTE2VTx?= M2PnNmlvcGmkaYTpc44hd2ZiU2TBWFEh[W6mIGPURXQ{KHCqb4PwbI9zgWyjdHnvckB{cWewaX\pZ4FvfGy7 NEX0NWszOzl2MUizNi=>
SNU423 MUjGeY5kfGmxbjDBd5NigQ>? MWq1NEDPxE1? M1;ibFI1KGh? NHTsO3RFVVOR M4fuUmlvcGmkaYTpc44hd2ZiU2TBWFEh[W6mIGPURXQ{KHCqb4PwbI9zgWyjdHnvckB{cWewaX\pZ4FvfGy7 NXHkUoRPOjN7NEG4N|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 slightly soluble or insoluble
Ethanol slightly soluble or insoluble
In vivo Add solvents individually and in order:
2% DMSO+30% PEG 300+ddH2O
5mg/mL

* 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