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 236 Publications

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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 MXnLbY5ie2ViQYPzZZk> MkL2NlAhdWmw NED2SHNFVVOR MkjQTY5pcWKrdHnvckBw\iCMQVuyJIlvKGi3bXHuJHRHOSClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGVRVy2rbnT1Z4VlKFOWQWS1JJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMUNOwG0> NEDKPWozOjZ7OEC4OC=>
TF1 MlTVT4lv[XOnIFHzd4F6 NV3ld5NQOjBibXnu M{XHeGROW09? MnG1TY5pcWKrdHnvckBw\iCMQVuxJIlvKGi3bXHuJHRHOSClZXzsd{Bie3Onc4Pl[EBieyCrbnjpZol1cW:wIH;mJGlNPi2rbnT1Z4VlKFOWQWSzJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yMkVOwG0> MkfYNlI3QThyOES=
Human T cell NHHXU4pMcW6jc3WgRZN{[Xl? MnL1TY5pcWKrdHnvckBw\iCMQVuzM|EhcW5iaIXtZY4hXCClZXzsd{BmgHC{ZYPzbY5oKEOGMzDhd5Nme3OnZDDhd{BqdmirYnn0bY9vKG:oIFnMNk1{fGmvdXzheIVlKFOWQWS1ZUBxcG:|cHjvdplt[XSrb36ge4l1cCCLQ{WwJI9nKDBwMEKz{txO NXjXNYllOjN3NEC2OFg>
Human monocyte NFPvfY1McW6jc3WgRZN{[Xl? M{nBZWlvcGmkaYTpc44hd2ZiSlHLNkBqdiCqdX3hckBud26xY4n0[ZMh\XiycnXzd4lv\yCFREG0JIF{e2W|c3XkJIF{KGmwaHnibZRqd25ib3[gS20uS1OILYP0bY12dGG2ZXSgV3RCXDWjIIDoc5NxcG:{eXzheIlwdiC5aYToJGlEPTBib3[gNE4xOjcQvF2= M{HDSVI{PTRyNkS4
Human monocyte MkfQT4lv[XOnIFHzd4F6 M4nwS2lvcGmkaYTpc44hd2ZiSlHLNk8yKGmwIHj1cYFvKG2xbn;jfZRmeyCneIDy[ZN{cW6pIFPENVQh[XO|ZYPz[YQh[XNiaX7obYJqfGmxbjDv[kBKTk6pYX3tZU1{fGmvdXzheIVlKFOWQWSxJJBpd3OyaH;yfYxifGmxbjD3bZRpKEmFNUCgc4YhOC5yM{JOwG0> NHLLNHMzOzV2ME[0PC=>
HEL MYDDfZRwfG:6aXOgRZN{[Xl? M3zOdFUh|ryP M4mzbVQ5KGh? NUPJVHZyS3m2b4TvfIlkKGmwZHX4QVEzNjJn NXvHW4NwOjV7M{GzOFk>
SET-2 M2\0bWN6fG:2b4jpZ{BCe3OjeR?= NWPFNIk6PSEQvF2= MknCOFghcA>? NXzacm5ZS3m2b4TvfIlkKGmwZHX4QVE5Njdn MUKyOVk{OTN2OR?=
HT93A Mo[zS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? MlG1N|IxKG6P MojFOUBl MorpSG1UVw>? MV;Jcohq[mm2aX;uJI9nKEeFUz3GJIlv\HWlZXSg[5JidnWub3P5eIlkKGSrZn\ldoVvfGmjdHnvci=> NYjw[XlrOjV6MEW5OlI>
CMK MVLHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NW[wWJlOUW6qaXLpeIlwdiCxZjDDUWsh[2G{conpcochfGinIFrBT|NCPTd{VjDteZRifGmxbjDj[YxtKHC{b3zp[oVz[XSrb36= NYrEW3d[OjV|NUKxNlQ>
CMK MYnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MoHrTY5pcWKrdHnvckBw\iCFTVugZ4FzenmrbnegeIhmKEqDS{PBOlNFKG23dHH0bY9vKGOnbHygdJJwdGmoZYLheIlwdiC5aYToJGlEPTBib3[gNE4yPjNizszN NVztS241OjV|NUKxNlQ>
CMK M2PQRWdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 MXLJcohq[mm2aX;uJI9nKEOPSzDjZZJzgWmwZzD0bIUhX1RiSlHLJINmdGxicILvcIln\XKjdHnvckB4cXSqIFnDOVAhd2ZiMD6wO|Uh|ryP MkLVNlU{PTJzMkS=
NCI-H460 M1roPGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NHjWcFJFVVOR NI\QSm9KSzVyPUCuNVMh|ryP NUDwb5lJOjV{MUO2O|A>
NCI-H358 NH\1UHNIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NYSwSplTTE2VTx?= NEW5dpBKSzVyPUCuNUDPxE1? MV[yOVIyOzZ5MB?=
A549 NH7nUIdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= MXrEUXNQ MVfJR|UxRTBwMESg{txO MYmyOVIyOzZ5MB?=
A549/DDP Mn[0S5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NFjTS21FVVOR MoDrTWM2OD1yLkKyJO69VQ>? MVuyOVIyOzZ5MB?=
NCI-H1299 M2fs[Wdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NFP1OJdFVVOR MXnJR|UxRTBwMkig{txO M13KZ|I2OjF|Nkew
NCI-H2347 NUTkWYxwT3Kxd4ToJGlvcGmkaYTpc44hSXO|YYm= NUftUnV7TE2VTx?= MVLJR|UxRTBwMUeg{txO NVHCeFVKOjV{MUO2O|A>
A549/DDP MVfGeY5kfGmxbjDBd5NigQ>? NX7DWJZlOzBibl2= M1;N[FQ5KGh? NVXYR|A{TE2VTx?= MkLoSI94di2{ZXf1cIF1cW:wIH;mJHNVSVR|IIDoc5NxcG:{eXzheIlwdg>? MlS5NlUzOTN4N{C=
NCI-H1299 NVXGeGpvTnWwY4Tpc44hSXO|YYm= MUmzNEBvVQ>? MYm0PEBp NH7vZW5FVVOR M2XycWRwf25vcnXneYxifGmxbjDv[kBUXEGWMzDwbI9{eGixconsZZRqd25? NI\Zdo8zPTJzM{[3NC=>
NCI-H2347 M37qemZ2dmO2aX;uJGF{e2G7 MYWzNEBvVQ>? MoDIOFghcA>? NGTMfHFFVVOR MlS2SIVkemWjc3WgbY4hSmOuMjDlfJBz\XO|aX;u NXfwb3JmOjV{MUO2O|A>
A549/DDP NHLJeYdCeG:ydH;zbZMhSXO|YYm= NG\qfIY{OCCwTR?= M{nKe|Q5KGh? M2i3fWROW09? MoTxTY5lfWO2aX;uJI9nKGGyb4D0c5Nqew>? NW\kV5FNOjV{MUO2O|A>
NCI-H1299 NYjtNIpGSXCxcITvd4l{KEG|c3H5 M3HVcFMxKG6P M2DTUlQ5KGh? NXvYO4dLTE2VTx?= MUjJcoR2[3Srb36gc4Yh[XCxcITvd4l{ MmPWNlUzOTN4N{C=
NCI-H2347 NUXE[Zd4SXCxcITvd4l{KEG|c3H5 M1:yd|MxKG6P NHvoT5c1QCCq Mn[0SG1UVw>? MXHJcoR2[3Srb36gc4Yh[XCxcITvd4l{ NVvibVVGOjV{MUO2O|A>
Hep3B MYnGeY5kfGmxbjDBd5NigQ>? NH7meWUyKM7:TR?= NYjnW45COTZiaB?= NGDsZ21FVVOR NVL2[FBlUW2yYXny[ZMhfGinIHPhdIFkcXS7IH;mJGlJS0FvYYPzc4Nq[XSnZDDndFE{OCCvdYThcpR{KHSxIHHjeIl3\SCVVFHUN{B4cXSqIFnDOVAhd2ZifkWwJO69VQ>? MV[yOFUxOTZ6OR?=
HepG2 NVjVbHc{TnWwY4Tpc44hSXO|YYm= M2q3[VEh|ryP NWLaUJliOTZiaB?= NFHxUnBFVVOR MVHJcZBicXKnczD0bIUh[2GyYXPpeJkhd2ZiSVjDRU1ie3OxY3nheIVlKGeyMUOwJI12fGGwdIOgeI8he2mpbnHsJJRwKFOWQWSz M1zjOVI1PTBzNki5
Huh7 M3f5fWZ2dmO2aX;uJGF{e2G7 MXKxJO69VQ>? MWmxOkBp MlfSSG1UVw>? NIP4SoZKdXCjaYLld{B1cGViY3HwZYNqfHlib3[gTWhESS2jc4PvZ4lifGWmIHfwNVMxKG23dHHueJMhfG9ic3nncoFtKHSxIGPURXQ{ NGLqNJEzPDVyMU[4PS=>
BaF3 NHW0eWFMcW6jc3WgRZN{[Xl? NXPuOY9JQDBibl2= MXq2JIg> M1\DeGROW09? M4m0dXJm\HWlZYOgeIhmKHCqb4PwbI9zgWyjdHnvckBw\sLiU2TBWFUhcW5iSlHLNnY3OTeILX31eIF1\WRiQlHGN{1GWE:UIHPlcIw> NITBcnUzPDJ|N{e5NS=>
DLD-1 NVrXUVRjU2mwYYPlJGF{e2G7 MoqyNlUh|ryP MXu0PEBp NXzHZVhuTE2VTx?= MWDJcohq[mm2aX;uJI9nKEqDS{GgdIhwe3Cqb4L5cIF1cW:w MV6yOFA2ODV3MB?=
RKO MkPhT4lv[XOnIFHzd4F6 NEe2flUzPSEQvF2= NYq3fVVUPDhiaB?= MlvVSG1UVw>? MV\Jcohq[mm2aX;uJI9nKEqDS{GgdIhwe3Cqb4L5cIF1cW:w NEXY[IQzPDB3MEW1NC=>
DLD-1 NYP6XVJLU2mwYYPlJGF{e2G7 MoTuNlUh|ryP NV24TVZzPDhiaB?= NHeyN5RFVVOR MVnJcohq[mm2aX;uJI9nKEqDS{KgdIhwe3Cqb4L5cIF1cW:w NVrobpJkOjRyNUC1OVA>
RKO MVHLbY5ie2ViQYPzZZk> MmfPNlUh|ryP Mn3aOFghcA>? MVrEUXNQ NX\XOW9s\G:nczDuc5QhcW6qaXLpeEBLSUtzIIDoc5NxcG:{eXzheIlwdg>? MoH4NlQxPTB3NUC=
DLD-1 M4jrNGdzd3e2aDDJcohq[mm2aX;uJGF{e2G7 NGDE[mI2OCEQvF2= MlLoOFghcA>? MnHQSG1UVw>? MVvJR|UxRTF3LkWxJO69VQ>? MVyyOFA2ODV3MB?=
RKO NGfDZpdIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= NUTCUo8{PTBizszN M1LwdFQ5KGh? M4\ifWROW09? MlLwTWM2OD1zND63OkDPxE1? NHT6T24zPDB3MEW1NC=>
DLD-1 NGm2b|ZCeG:ydH;zbZMhSXO|YYm= M{HW[|I2KM7:TR?= M2rkS|Q5KGh? NUDiWHhKTE2VTx?= MoTsTY5lfWOnczDhdI9xfG:|aYOgZpkh[WO2aY\heIlv\yClYYPwZZNmKDN? M4DTSFI1ODVyNUWw
RKO NFj0PHdCeG:ydH;zbZMhSXO|YYm= M{jwfFI2KM7:TR?= MoPkOFghcA>? MU\EUXNQ MYjJcoR2[2W|IHHwc5B1d3OrczDifUBi[3SrdnH0bY5oKGOjc4Dhd4UhOw>? M{TGZ|I1ODVyNUWw
HuH7 MWXHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? Mny3OVAh|ryP M4\WOVQ5KGh? M3rh[2ROW09? MmG1QlgzLSC{ZXT1Z5Rqd25? M2jCclI{QTRzOEOy
SNU182 MknoS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? M1nzcFUxKM7:TR?= NYCye5hZPDhiaB?= NETxTpNFVVOR NWPkTXBWRjZ2JTDy[YR2[3Srb36= NXXrcFJZOjN7NEG4N|I>
SNU423 MYrHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? MXK1NEDPxE1? MlfTOFghcA>? NX\oSnNRTE2VTx?= NGf0O|U,QDFnIILl[JVkfGmxbh?= MUWyN|k1OTh|Mh?=
HuH7 MoC5SpVv[3Srb36gRZN{[Xl? NIDkbHI2OCEQvF2= NWjrNIhEOjRiaB?= NWfldlhmTE2VTx?= NX72c2pwUW6qaXLpeIlwdiCxZjDTWGFVOSCjbnSgV3RCXDNicHjvd5Bpd3K7bHH0bY9vKHOrZ37p[olk[W62bIm= M3\GOFI{QTRzOEOy
SNU182 M1fXcmZ2dmO2aX;uJGF{e2G7 M4P2WFUxKM7:TR?= M{nG[VI1KGh? NVXmNYM{TE2VTx?= NF3tT4dKdmirYnn0bY9vKG:oIGPURXQyKGGwZDDTWGFVOyCyaH;zdIhwenmuYYTpc44he2mpbnnmbYNidnSueR?= MXeyN|k1OTh|Mh?=
SNU423 NFzDSnVHfW6ldHnvckBCe3OjeR?= Mo\6OVAh|ryP MVuyOEBp MmXTSG1UVw>? Ml;ITY5pcWKrdHnvckBw\iCVVFHUNUBidmRiU2TBWFMheGixc4Doc5J6dGG2aX;uJJNq\26rZnnjZY51dHl? NGjkTIczOzl2MUizNi=>

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
cleaved PARP / cleaved caspase3; 

PubMed: 29849942     


OVCAR-8 and MDAH 2774 cells were incubated with various concentrations of ruxolitinib for 48 h. Apoptosis was determined by using cleaved poly-ADP ribose polymerase (PARP) and cleaved caspase-3 by Western blot.

p-JAK2 / p-AKT / p-MAPK / Bcl-xl / MCL-1; 

PubMed: 29849942     


OVCAR-8 and MDAH2774 cells were treated with ruxolitinib (20 μM), paclitaxel (10 nM) or the combination for 24 h. Whole cells were collected and determined for the change of STAT3, AKT and ERK pathways and expression of BCL-XL and MCL-1 by Western blot.

c-Myc / c-Jun / Cyclin B / Cyclin D / Bcl-2 / HIF-1α; 

PubMed: 30930994     


Effects of ruxolitinib on the expression of downstream target genes of the JAK-STAT pathway. The protein levels of c-Myc, c-Jun, Cyclin B1, Cyclin D1, Bcl-2 and HIF-1α were determined in MCF-7 and TAMR-MCF-7 cells 24 h following ruxolitinib treatment (0.1-10 μM). 

p-STAT3; 

PubMed: 29849942     


Dose-dependent inhibition of STAT3 phosphorylation. Human ovarian cancer cells, OVCAR-8, MDAH2774, and SKOV3, were treated with the indicated concentrations of ruxolitinib for 24 h. Phosphorylation of STAT3 was analyzed by Western blot. 

29849942 30930994
Growth inhibition assay
Cell viability; 

PubMed: 29849942     


Dose dependent inhibition of cell viability. Human ovarian cancer cell lines were treated with the indicated concentrations of ruxolitinib. Cell viability was determined 72 h later. The IC50 was determined by the Chou-Talalay method. *P<0.05; ***P<0.0005, ruxolitinib vs control in OVCAR-8 cells; #P<0.05; ##P<0.005; ###P<0.0005, ruxolitinib vs control in SKOV-3 cells; ^^P<0.005; ^^^P<0.0005, ruxolitinib vs control in MDAH2774 cells.

Cell apoptosis; 

PubMed: 29849942     


OVCAR-8 and MDAH 2774 cells were incubated with various concentrations of ruxolitinib for 48 h. Apoptosis was determined by flow cytometry using annexin V and PI staining.

Cell proliferation; 

PubMed: 29515770     


Cells were plated into 48 well plates and cell growth was measured every 48 hours via MTS assay following ruxolitinib treatment (0, 1, 10 and 100 uM) in L-428 (left) and HDLM-2 (middle) HL cells, and Karpas-1106P PMBL cells (right).

29849942 29515770
Immunofluorescence
α-tubulin; 

PubMed: 26356819     


Confocal analysis of HEL cells, treated or not with different concentration of ruxolitinib (100 and 300 nM), displaying α-Tubulin (green) and DAPI (blue) staining; MERGE shows the overlapped images. Scale bars are shown in the figure (10 μm). Note more diffuse microtubule networks in ruxolutinib-treated cells.

26356819
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]
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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]
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  • 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]
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  • 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)
Ethanol 61 mg/mL (199.1 mM)
Water Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+ddH2O
For best results, use promptly after mixing.
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 interventions Conditions Sponsor/Collaborators Start Date Phases
NCT03774082 Not yet recruiting Drug: INC424 Graft vs Host Disease Novartis Pharmaceuticals|Novartis October 8 2019 Phase 2
NCT04057573 Not yet recruiting Drug: Ruxolitinib cream|Drug: Vehicle Non-segmental Vitiligo Incyte Corporation September 10 2019 Phase 3
NCT04052425 Not yet recruiting Drug: Ruxolitinib cream|Drug: Vehicle Non-segmental Vitiligo Incyte Corporation September 10 2019 Phase 3

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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Frequently Asked Questions

  • Question 1:

    What is the difference between S2902 and S1378 which seem to have same structure formula according to the product information?

  • Answer:

    These two chemicals are the two different chiral forms of Ruxolitinib. S2902 S-Ruxolitinib is the S form and S1378 Ruxolitinib is the D form. One of the carbon atoms in this molecule is asymmetric, making the two molecules mirror images of each other. The biological activities of these two molecules can be very different because of the confirmation differences.

  • Question 2:

    How about the half-life of the compound (Ruxolitinib)? How long is the duration of the inhibitory effect on JAK-STAT signaling?

  • Answer:

    The half-life of this compound in body is about 2~3 hours according to previous study. Generally, it is longer in vitro culture medium than in vivo. In paper, Ruxolitinib was also used for 24hours. http://www.bloodjournal.org/cgi/pmidlookup?view=long&pmid=24711661.

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID