RO4929097

For research use only.

Catalog No.S1575 Synonyms: RG-4733

93 publications

RO4929097 Chemical Structure

CAS No. 847925-91-1

RO4929097 (RG-4733) is a γ secretase inhibitor with IC50 of 4 nM in a cell-free assay, inhibiting cellular processing of Aβ40 and Notch with EC50 of 14 nM and 5 nM, respectively. Phase 2.

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Selleck's RO4929097 has been cited by 93 publications

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Biological Activity

Description RO4929097 (RG-4733) is a γ secretase inhibitor with IC50 of 4 nM in a cell-free assay, inhibiting cellular processing of Aβ40 and Notch with EC50 of 14 nM and 5 nM, respectively. Phase 2.
Targets
γ secretase [1]
(Cell-free assay)
Notch [1]
(Cell-free assay)
Aβ40 [1]
(Cell-free assay)
4 nM 5 nM 14 nM
In vitro

RO4929097 decreases the amount of Aβ peptides secreted into the culture medium in HEK293 cells with EC50 of 14 nM. RO4929097 strongly inhibits Notch processing with EC50 of 5 nM in the Notch cell-based reporter assay. The potency of RO4929097 in cell-free and cellular assays is in the low nanomolar range with >100-fold selectivity observed with respect to 75 other proteins of various types including receptors, ion channels, and enzymes (CEREP panel). After 5 days of treatment, RO4929097 reduces the production of ICN in the human NSCLC A549 cells inducing a flattened and less transformed tumor cell phenotype in tissue culture. [1] RO4929097 blocks Notch processing in human non-small cell lung carcinoma cells and decreases expression of the Notch transcriptional target gene Hes1. Treatment with RO4929097 reveals a two- to threefold decrease in the expression of direct Notch target genes, Hes1, Hey1, and Heyl in SUM149 and a 3.5- to eightfold decrease in expression in SUM190 cells. RO4929097 modestly inhibits the growth of SUM149 cells in a dose-dependent manner. At a concentration of 1 μM of RO4929097, growth inhibition is 20 % for SUM149 and 10 % for SUM190 cells, relative to vehicle-treated controls. RO4929097 decreases the production of inflammatory cytokines by T-cells. Furthermore, with RO4929097 treatment, there is a shift in favor of TH2 over TH1 cytokines. In addition, T-cell activation induced IL-6 production would be increased with RO4929097. [2] Upon RO4929097 treatment, the selected melanoma cell lines reveals downregulation of NOTCH downstream effector HES1. A decrease in the amount of melanospheres formed upon RO4929097 treatment in primary melanoma cell lines is detected. [3]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
U87 MVLD[YxtKF[rYXLpcIl1gSCDc4PhfS=> NHvwXGU{OCEQvF2= MYC0JIQ> NITM[o9l\WO{ZXHz[ZMh[2WubDD2bYFjcWyrdIm= NW\Yc5U5RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS0PVU6ODdpPkK0OFk2QTB5PD;hQi=>
U87 R1 M4jTXmNmdGxiVnnhZoltcXS7IFHzd4F6 NHf3emU{OCEQvF2= MnrhOEBl NVnkOJpz\GWlcnXhd4V{KGOnbHygeoli[mmuaYT5 NGHudmQ9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{NES5OVkxPyd-MkS0PVU6ODd:L3G+
MGG4 MXjD[YxtKF[rYXLpcIl1gSCDc4PhfS=> MkDLN|Ah|ryP NVm3[I1[PCCm NUTWfY9I\GWlcnXhd4V{KGOnbHygeoli[mmuaYT5 Mn\6QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjR2OUW5NFcoRjJ2NEm1PVA4RC:jPh?=
MGG6 MkTXR4VtdCCYaXHibYxqfHliQYPzZZk> M17MXVMxKM7:TR?= NF3TWJo1KGR? MXrk[YNz\WG|ZYOgZ4VtdCC4aXHibYxqfHl? NWDMSoZGRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS0PVU6ODdpPkK0OFk2QTB5PD;hQi=>
MGG8 M3HuS2NmdGxiVnnhZoltcXS7IFHzd4F6 MXyzNEDPxE1? M{fWRVQh\A>? NIL2W5Zl\WO{ZXHz[ZMh[2WubDD2bYFjcWyrdIm= MYC8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zPDR7NUmwO{c,OjR2OUW5NFc9N2F-
MGG23 Mn\0R4VtdCCYaXHibYxqfHliQYPzZZk> MWWzNEDPxE1? NFjEPFQ1KGR? NYO2S4ts\GWlcnXhd4V{KGOnbHygeoli[mmuaYT5 NVz5cVIxRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkS0PVU6ODdpPkK0OFk2QTB5PD;hQi=>
SUM149 Mn;ZR4VtdCCYaXHibYxqfHliQYPzZZk> M3TBd|AuOTBizszN NF;WcVI4OiCq M4rnWYlvcGmkaYTzJINmdGxiZ4Lve5RpKGSxc3Wg[IVx\W6mZX70cJk> NGO1N|k9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MkW0O|ExQSd-MkK1OFcyODl:L3G+
Sum190 MUPD[YxtKF[rYXLpcIl1gSCDc4PhfS=> MlHyNE0yOCEQvF2= NFHURmQ4OiCq NFznVm5qdmirYnn0d{Bk\WyuIHfyc5d1cCCmb4PlJIRmeGWwZHXueIx6 NVvzOGZ5RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkK1OFcyODlpPkKyOVQ4OTB7PD;hQi=>
WM35 M2HJU2Z2dmO2aX;uJGF{e2G7 NYLLcXlROTBidV2= M{jxOVI1KGh? NEnie|VFVVOR NGDhPG9l\WO{ZXHz[ZMhfGinIHzleoVteyCxZjDOU3REUCCmb4fud5Rz\WGvIIThdodmfCCKRWOx NWKwellQRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG5PFA1ODhpPkKxPVgxPDB6PD;hQi=>
WM98.1 NFyyOoNHfW6ldHnvckBCe3OjeR?= MnG0NVAhfU1? MlLxNlQhcA>? NHPLZ2xFVVOR NVf5Z2g4\GWlcnXhd4V{KHSqZTDs[ZZmdHNib3[gUm9VS0hiZH;3cpN1emWjbTD0ZZJo\XRiSFXTNS=> M2S5UVxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzOUiwOFA5Lz5{MUm4NFQxQDxxYU6=
WM115 NYjycIpzTnWwY4Tpc44hSXO|YYm= MXmxNEB2VQ>? NHf3O4UzPCCq NGewZYRFVVOR MULk[YNz\WG|ZYOgeIhmKGyndnXsd{Bw\iCQT2TDTEBld3ewc4Ty[YFuKHSjcnfleEBJTVNz NXT5ZmxTRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG5PFA1ODhpPkKxPVgxPDB6PD;hQi=>
WM983A Mor4SpVv[3Srb36gRZN{[Xl? NF7FS3cyOCC3TR?= MnnVNlQhcA>? M1XFdmROW09? NF;vZ4Nl\WO{ZXHz[ZMhfGinIHzleoVteyCxZjDOU3REUCCmb4fud5Rz\WGvIIThdodmfCCKRWOx M2HKV|xiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzOUiwOFA5Lz5{MUm4NFQxQDxxYU6=
WM3248  NYLiVG9TTnWwY4Tpc44hSXO|YYm= NIjCZYoyOCC3TR?= M4nxPVI1KGh? MlzpSG1UVw>? NHzPPIVl\WO{ZXHz[ZMhfGinIHzleoVteyCxZjDOU3REUCCmb4fud5Rz\WGvIIThdodmfCCKRWOx NUX6eHJNRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG5PFA1ODhpPkKxPVgxPDB6PD;hQi=>
WM35 MmPlS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NVnwNXdpOTBidV2= NYfUWXFYOC1zODDk NYTzSVJRTE2VTx?= MnH0bY5pcWKrdIOgZ4VtdCCpcn;3eIghfGmvZTDk[ZBmdmSnboTsfS=> NGLKOGs9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{MUm4NFQxQCd-MkG5PFA1ODh:L3G+
WM98.1 NEHySWRIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= Mk\RNVAhfU1? NWP0OHF6OC1zODDk MmmzSG1UVw>? NX\ZR|FYcW6qaXLpeJMh[2WubDDndo94fGhidHnt[UBl\XCnbnTlcpRtgQ>? M3;GcFxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJzOUiwOFA5Lz5{MUm4NFQxQDxxYU6=
WM115 Mn\IS5Jwf3SqIFnubIljcXSrb36gRZN{[Xl? NWDkdoxXOTBidV2= MWOwMVE5KGR? Mm\MSG1UVw>? NHPwVW1qdmirYnn0d{Bk\WyuIHfyc5d1cCC2aX3lJIRmeGWwZHXueIx6 MlHTQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjF7OEC0NFgoRjJzOUiwOFA5RC:jPh?=
WM983A MYnHdo94fGhiSX7obYJqfGmxbjDBd5NigQ>? NW\jUppbOTBidV2= NHTsb5IxNTF6IHS= NUeyWnBFTE2VTx?= MWHpcohq[mm2czDj[YxtKGe{b4f0bEB1cW2nIHTldIVv\GWwdHz5 MU[8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zOTl6MESwPEc,OjF7OEC0NFg9N2F-
WM3248  NHvWWYJIem:5dHigTY5pcWKrdHnvckBCe3OjeR?= M33S[|ExKHWP NWjLfJRKOC1zODDk MnjOSG1UVw>? NX;LdHFqcW6qaXLpeJMh[2WubDDndo94fGhidHnt[UBl\XCnbnTlcpRtgQ>? NWTuTGxjRGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkG5PFA1ODhpPkKxPVgxPDB6PD;hQi=>
A673 MV;xTHRUKGG|c3H5 MnnoQIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjl2M{WxN|koRjJ7NEO1NVM6RC:jPh?=
DAOY NUDBOnR2eUiWUzDhd5NigQ>? NHX1[ZE9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OUSzOVE{QSd-Mkm0N|UyOzl:L3G+
BT-37 Mnr6dWhVWyCjc4PhfS=> NELqU5g9[SC2YYLn[ZQ:L1:kbHHub{chcHKnZk2nbJR1eHN8Lz;weYJu\WRwbnPibU5vdG1wbnnoModwfi9{OUSzOVE{QSd-Mkm0N|UyOzl:L3G+
BT-12 M13afJFJXFNiYYPzZZk> M2fEUlxiKHSjcnfleF0oZ2KuYX7rK{BpemWoPTfoeJRxezpxL4D1Zo1m\C6wY3LpMo5tdS6waXiu[493NzJ7NEO1NVM6Lz5{OUSzOVE{QTxxYU6=
OHS-50 NIXYO3JyUFSVIHHzd4F6 MWS8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTR|NUGzPUc,Ojl2M{WxN|k9N2F-
SJ-GBM2 NFv1Z5dyUFSVIHHzd4F6 NXXNelc4RGFidHHy[4V1RSehYnzhcosoKGi{ZX[9K4h1fHC|Oj:vdJVjdWWmLn7jZokvdmyvLn7pbE5od3ZxMkm0N|UyOzlpPkK5OFM2OTN7PD;hQi=>
SK-N-MC M3v3NpFJXFNiYYPzZZk> Mm[4QIEhfGG{Z3X0QUdg[myjbnunJIhz\WZ;J3j0eJB{Qi9xcIXicYVlNm6lYnmucoxuNm6raD7nc5YwOjl2M{WxN|koRjJ7NEO1NVM6RC:jPh?=
NB-EBc1 NEXGOG9yUFSVIHHzd4F6 MVO8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTR|NUGzPUc,Ojl2M{WxN|k9N2F-
LAN-5 NGDsdJJyUFSVIHHzd4F6 MYi8ZUB1[XKpZYS9K39jdGGwazegbJJm\j1paIT0dJM7Ny:ydXLt[YQvdmOkaT7ucI0vdmmqLnfvek8zQTR|NUGzPUc,Ojl2M{WxN|k9N2F-

... Click to View More Cell Line Experimental Data

Assay
Methods Test Index PMID
Western blot
Hey1; 

PubMed: 29899322     


HeLa and Caski cells treated with RO4929097 and the DMSO control showed downregulation of the Hey1 protein as determined by Western blot analysis;

Snail / N-cadherin / Twist / E-cadherin; 

PubMed: 29899322     


Effect of inhibiting the Notch signaling pathway on the protein expression levels of EMT-associated proteins Snail, N-cadherin, Twist, and E-cadherin in HeLa and Caski cells. 

Akt / p-Akt / Notch / IGF1R / FBXW7 ; 

PubMed: 28507201     


Western blots showing expression of p-Akt, Akt, Notch, IGF1R, and FBXW7 in HCC827/ER cells treated with MK-2206 (an Akt inhibitor) or RO4929097 (a Notch inhibitor). GAPDH served as an internal control. 

NICD / Hes1 / Hes3 / Hes5; 

PubMed: 24038660     


A panel of GICs was treated with the indicated doses of DAPT, BMS-708163 and RO4929097 for 48 hours. γ Secretase inhibitors inhibited expression of NICD, Hes1, Hes3 and Hes5 in a dose-dependent manner.

29899322 28507201 24038660
Growth inhibition assay
Cell viability; 

PubMed: 30669546     


A panel of GIC lines was treated with various concentrations of the RO4929097. Cells were treated in triplicate wells for 72 h, and cell viability was assessed with the CellTiter-Blue assay. Cell viability in the vehicle control was considered to be 100%; 

30669546
In vivo Oral injection of 3 to 60 mg/kg RO4929097 once daily or twice daily to nude mice bearing A549 NSCLC xenografts for either 7, 14, or 21 days of a 21-day schedule results in significant tumor growth inhibition compared with vehicle-treated animals. The tumor growth inhibition values ranges from 66% to 91%. When mice are treated with 60 mg/kg RO4929097 twice daily with the 7+/14- schedule, treatment initially arouses regression of established A549 tumors. At the end of the 21-day cycle (day 47), tumor growth prevention is still 91% compared with vehicle control mice. Inhibition of tumor growth remains prolonged and sustained up to 34 days post-treatment (day 67). On day 67, these mice are retreated with the same dose of RO4929097 for a second cycle (7 days) until day 74. Importantly, the antitumor effects are sustained after dosing is completed. [1] RO4929097 leads to reduced expression of genes associated with angiogenesis in A549 xenograft model. In contrast, the RO4929097-resistant H460a xenograft displays little change in expression of these genes, underscoring the in vivo anti-angiogenesis mechanism of action of RO4929097.[2] For IL6 and IL8 overexpressing tumors, RO4929097 no longer impacts angiogenesis or the infiltration of tumor associated fibroblasts. [4]

Protocol

Kinase Assay:[5]
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In vitro potency assays:

After RO4929097 is used, the Aβ peptides are measured by ECL assays using a variety of anti-Aβ antibodies and an Origen 1.5 Analyzer. The 4G8 murine mAb binds an epitope in the Aβ peptide (within amino acids 18–21) that is immediately distal to the α-secretase cleavage site. The G2–10 murine mAb binds the C terminus that is exposed after γ-secretase-mediated cleavage to generate amino acid 40 of the Aβ40 peptide. The FCA3542 rabbit antibody binds the C terminus that is exposed after γ-secretase-mediated cleavage to generate amino acid 42 of the Aβ42 peptide. The 4G8 mAb is biotinylated with biotin-LC-sulfo-N-hydroxysuccinimide-ester. The G2–10 and FCA3542 antibodies are ruthenylated with TAG-N-hydroxysuccinimide ester. Aβ(x-40) is detected with biotinylated 4G8 and ruthenylated G2–10. Aβ(x-42) is detected with biotinylated 4G8 and ruthenylated FCA3542.
Cell Research:[3]
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  • Cell lines: WM35 and WM98.1 cell lines
  • Concentrations: 10 μM
  • Incubation Time: DMSO
  • Method: Primary melanoma cell lines, including WM35 and WM98.1, are seeded at 2.5 × 103 cells per well on a 12-well dish in triplicate. The day after (day 0), the medium is replaced, and DMSO or 10 μM RO4929097 is added and changed every 3-4 days. At the indicated time points, cells are fixed in 10% formalin solution and stored in PBS at 4 °C. At day 18-24, all the plates are stained with crystal violet. After color elution with 10% acetic acid, optical density is read at 590 nm.
    (Only for Reference)
Animal Research:[1]
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  • Animal Models: Female nude mice bearing Calu-6 cells
  • Dosages: 3 to 60 mg/kg
  • Administration: Oral administration
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 93 mg/mL (198.12 mM)
Water Insoluble
Ethanol ''16 mg/mL
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
2% DMSO+30% PEG 300+5% Tween+ddH2O
For best results, use promptly after mixing.
10mg/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 469.4
Formula

C22H20F5N3O3

CAS No. 847925-91-1
Storage powder
in solvent
Synonyms RG-4733
Smiles CC(C)(C(=O)NCC(C(F)(F)F)(F)F)C(=O)NC1C2=CC=CC=C2C3=CC=CC=C3NC1=O

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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.

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Frequently Asked Questions

  • Question 1:

    How about the half-life of RO4929097(S1575)?

  • Answer:

    For S1575, the half-life is about 20 hours based on the following paper: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3869895/

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID