PF-00562271 Besylate

Catalog No.S2672 Synonyms: PF-562271 Besylate

PF-00562271 Besylate Chemical Structure

Molecular Weight(MW): 665.66

PF-00562271 Besylate is the benzenesulfonate salt of PF-562271, which is a potent, ATP-competitive, reversible inhibitor of FAK with IC50 of 1.5 nM, ~10-fold less potent for Pyk2 than FAK and >100-fold selectivity against other protein kinases, except for some CDKs. Phase 1.

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  • Mol Ther 2012 20(5), 972-83. PF-00562271 Besylate purchased from Selleck.

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Biological Activity

Description PF-00562271 Besylate is the benzenesulfonate salt of PF-562271, which is a potent, ATP-competitive, reversible inhibitor of FAK with IC50 of 1.5 nM, ~10-fold less potent for Pyk2 than FAK and >100-fold selectivity against other protein kinases, except for some CDKs. Phase 1.
Targets
FAK [1]
(Cell-free assay)
PYK2 [1]
(Cell-free assay)
CDK2/CyclinE [1]
(Cell-free assay)
CDK3/CyclinE [1]
(Cell-free assay)
CDK1/CyclinB [1]
(Cell-free assay)
1.5 nM 13 nM 30 nM 47 nM 58 nM
In vitro

PF-562271 Besylate shows the selective inhibitory effects on FAK and Pyk2 tyrosine kinase activity with IC50 of 1.5 nM and 14 nM, respectively. And in cell-based assays, the IC50 of PF-562271 is shown to be 5 nM for FAK, which is more selective compared to other kinase targets. [1] In 2 dimensional (2D) cultures, PF-562271 results in a dose-dependent cell proliferation inhibition in FAK WT, FAK−/− and FAK kinase-deficient (KD) cells with IC50 of 3.3 μM, 2.08 μM and 2.01 μM, respectively. [2]

Cell Data
Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID
MV-4-11 cell Mm\vS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NIfGOnJKdmirYnn0bY9vKG:oIHj1cYFvKE2YLUStNVEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0xNjJ5Nk[g{txO M3KzOXNCVkeHUh?=
human SW982 cell MlTiS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NWDIWXhnUW6qaXLpeIlwdiCxZjDoeY1idiCVV{m4NkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvOzJ6MjFOwG0> NXf3Wm5FW0GQR1XS
human KM12 cell NVX6O49zT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MlrpTY5pcWKrdHnvckBw\iCqdX3hckBMVTF{IHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO|YYmsJGlEPTB;MD6zPFU2PyEQvF2= NHPNUJVUSU6JRWK=
human COLO-205 cell M33vZmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MlPFTY5pcWKrdHnvckBw\iCqdX3hckBEV0yRLUKwOUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvPDh4NUig{txO M3K5S3NCVkeHUh?=
human COLO-829 cell M1nGNWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MoWzTY5pcWKrdHnvckBw\iCqdX3hckBEV0yRLUiyPUBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvPzZzN{[g{txO NH7PfmlUSU6JRWK=
human MG-63 cell MWDHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? Mo\FTY5pcWKrdHnvckBw\iCqdX3hckBOTy14MzDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUCuPFA3OzdizszN NV\YS3Z2W0GQR1XS
human IGROV-1 cell NVy1XXV4T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NWnUblFXUW6qaXLpeIlwdiCxZjDoeY1idiCLR2LPWk0yKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVA:OC56MUCzPEDPxE1? M2jU[XNCVkeHUh?=
human NCI-H650 cell M1zXTmdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NYrvcHFiUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFY2OCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTBwOEOxOVQh|ryP NIj5c|FUSU6JRWK=
human RT-112 cell M2HjTGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 NHPMUGVKdmirYnn0bY9vKG:oIHj1cYFvKFKWLUGxNkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVAvQTh2NjFOwG0> NXPqN3ZkW0GQR1XS
human BCPAP cell M3LNPGdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 Ml3OTY5pcWKrdHnvckBw\iCqdX3hckBDS1CDUDDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNFEzQDhizszN NGHueGhUSU6JRWK=
ALL-PO cell MXfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MofKTY5pcWKrdHnvckBw\iCqdX3hckBCVExvUF:gZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlAyPTh2IN88US=> NGj0d3BUSU6JRWK=
human KYSE-270 cell NVjyUVNKT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MnLNTY5pcWKrdHnvckBw\iCqdX3hckBMYVOHLUK3NEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvODR5MUSg{txO NXXz[I9wW0GQR1XS
human 8305C cell M{LzOWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MofBTY5pcWKrdHnvckBw\iCqdX3hckA5OzB3QzDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNFk6ODRizszN NXzFS|NWW0GQR1XS
NCI-H810 cell M3K3Z2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MmS2TY5pcWKrdHnvckBw\iCqdX3hckBPS0lvSEixNEBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvOTB5N{[g{txO MlXuV2FPT0WU
human CAL-33 cell NF3xTI1Iem:5dHigbY5pcWKrdHnvckBie3OjeR?= MXHJcohq[mm2aX;uJI9nKGi3bXHuJGNCVC1|MzDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNVI6OzhizszN MmHLV2FPT0WU
human AN3-CA cell NXvzc3J4T3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= MYfJcohq[mm2aX;uJI9nKGi3bXHuJGFPOy2FQTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuNlE5PjdizszN M2m3WHNCVkeHUh?=
human NKM-1 cell MoLFS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? MVPJcohq[mm2aX;uJI9nKGi3bXHuJG5MVS1zIHPlcIwh\3Kxd4ToJIlvKGFiY3XscEB3cWGkaXzpeJkh[XO|YYmsJGlEPTB;MT6yO|UxPiEQvF2= MWHTRW5ITVJ?
human BPH-1 cell NX\OWoRHT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= M{fsfGlvcGmkaYTpc44hd2ZiaIXtZY4hSlCKLUGgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlI5PzZ4IN88US=> NYjQfFdqW0GQR1XS
human MES-SA cell NHn2TJVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= MUPJcohq[mm2aX;uJI9nKGi3bXHuJG1GWy2VQTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuN|A3QDJizszN M2SyU3NCVkeHUh?=
human CAL-62 cell M3;j[2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 M4qzOmlvcGmkaYTpc44hd2ZiaIXtZY4hS0GOLU[yJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU4{OTlyOTFOwG0> M4TCc3NCVkeHUh?=
human KYSE-150 cell M4jLO2dzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MUjJcohq[mm2aX;uJI9nKGi3bXHuJGt[W0VvMUWwJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU4{PTJ|NjFOwG0> M2rBN3NCVkeHUh?=
human SK-UT-1 cell MnfGS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NHn2V4VKdmirYnn0bY9vKG:oIHj1cYFvKFONLWXUMVEh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0yNjR2NkS3JO69VQ>? M3HucHNCVkeHUh?=
human HUTU-80 cell NIDofmVIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NXzVVVRoUW6qaXLpeIlwdiCxZjDoeY1idiCKVWTVMVgxKGOnbHyg[5Jwf3SqIHnuJIEh[2WubDD2bYFjcWyrdImgZZN{[XluIFnDOVA:OS52NEi4OkDPxE1? M1H4bnNCVkeHUh?=
human SIG-M5 cell MYDHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MUPJcohq[mm2aX;uJI9nKGi3bXHuJHNKTy2PNTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuOFg1QDdizszN NFfTe2ZUSU6JRWK=
human AGS cell MUfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NUnuS4VNUW6qaXLpeIlwdiCxZjDoeY1idiCDR2OgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlUzOTJ2IN88US=> M4DpWXNCVkeHUh?=
human ST486 cell MnfhS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? NUnOfGM5UW6qaXLpeIlwdiCxZjDoeY1idiCVVES4OkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvPTN{N{ig{txO MoDKV2FPT0WU
human HSC-2 cell Ml\lS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M3nxSGlvcGmkaYTpc44hd2ZiaIXtZY4hUFOFLUKgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlU{QTVizszN NUjuUHRTW0GQR1XS
human BC-1 cell MYfHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? M{HvSmlvcGmkaYTpc44hd2ZiaIXtZY4hSkNvMTDj[YxtKGe{b4f0bEBqdiCjIHPlcIwhfmmjYnnsbZR6KGG|c3H5MEBKSzVyPUGuOlE3PjRizszN MmK3V2FPT0WU
human CGTH-W-1 cell NFPzN5RIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NVTXT3NVUW6qaXLpeIlwdiCxZjDoeY1idiCFR2TIMXcuOSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTFwNkG2O|kh|ryP MkfpV2FPT0WU
human MZ1-PC cell NHzoOHRIem:5dHigbY5pcWKrdHnvckBie3OjeR?= M12zZWlvcGmkaYTpc44hd2ZiaIXtZY4hVVpzLWDDJINmdGxiZ4Lve5RpKGmwIHGgZ4VtdCC4aXHibYxqfHliYYPzZZktKEmFNUC9NU43OjNzMjFOwG0> NUK0S5UyW0GQR1XS
human SW1710 cell MYrHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? MoKxTY5pcWKrdHnvckBw\iCqdX3hckBUXzF5MUCgZ4VtdCCpcn;3eIghcW5iYTDj[YxtKH[rYXLpcIl1gSCjc4PhfUwhUUN3ME2xMlYzPjJ6IN88US=> MWDTRW5ITVJ?
human EW-13 cell NHP0R2JIem:5dHigbY5pcWKrdHnvckBie3OjeR?= NWXpb24xUW6qaXLpeIlwdiCxZjDoeY1idiCHVz2xN{Bk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVEvPjN2Nk[g{txO NFK2N3ZUSU6JRWK=
human U251 cell M{TEbWdzd3e2aDDpcohq[mm2aX;uJIF{e2G7 MYLJcohq[mm2aX;uJI9nKGi3bXHuJHUzPTFiY3XscEBoem:5dHigbY4h[SClZXzsJJZq[WKrbHn0fUBie3OjeTygTWM2OD1zLke0NFMyKM7:TR?= MVnTRW5ITVJ?
human NCI-H460 cell MXHHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NULDU2RtUW6qaXLpeIlwdiCxZjDoeY1idiCQQ1mtTFQ3OCClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTJwMES4N|kh|ryP M3TNSnNCVkeHUh?=
human DU-4475 cell MX7Hdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NI\JeZpKdmirYnn0bY9vKG:oIHj1cYFvKESXLUS0O|Uh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0zNjF2N{W5JO69VQ>? MnrHV2FPT0WU
human MFE-296 cell MoXKS5Jwf3SqIHnubIljcXSrb36gZZN{[Xl? M{PDVmlvcGmkaYTpc44hd2ZiaIXtZY4hVU[HLUK5OkBk\WyuIHfyc5d1cCCrbjDhJINmdGxidnnhZoltcXS7IHHzd4F6NCCLQ{WwQVIvPDd5OUKg{txO Ml\UV2FPT0WU
human DU-145 cell NU\ne45vT3Kxd4ToJIlvcGmkaYTpc44h[XO|YYm= NXTzOVVzUW6qaXLpeIlwdiCxZjDoeY1idiCGVT2xOFUh[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0zNjR7MUG4JO69VQ>? NWXnO4k5W0GQR1XS
human MDA-MB-231 cell MUTHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? M2nVU2lvcGmkaYTpc44hd2ZiaIXtZY4hVUSDLV3CMVI{OSClZXzsJIdzd3e2aDDpckBiKGOnbHygeoli[mmuaYT5JIF{e2G7LDDJR|UxRTJwNEm1O|Ih|ryP NWKyeI5oW0GQR1XS
human SNU-387 cell MUXHdo94fGhiaX7obYJqfGmxbjDhd5NigQ>? NIX6cmdKdmirYnn0bY9vKG:oIHj1cYFvKFOQVT2zPFch[2WubDDndo94fGhiaX6gZUBk\WyuII\pZYJqdGm2eTDhd5NigSxiSVO1NF0zNjV{OEKg{txO NECwfnFUSU6JRWK=

... Click to View More Cell Line Experimental Data

In vivo In several human s.c. xenograft models, PF-562271 exhibits dose-dependent tumor growth inhibition, and produces maximum tumor inhibition for PC-3M, BT474, BxPc3, and LoVo ranging from 78% to 94% inhibition at doses of 25 to 50 mg/kg twice daily, without weight loss, morbidity, or death. [1] PF-562271 (25 mg/kg by p.o.) leads to a significant decrease in tumor progression in both subcutaneous and bone metastasis PC3M-luc-C6 xenograft models. [3] In a Huh7.5 hepatocellular carcinoma xenograft model, combination therapy of sunitinib and PF-562271 targets angiogenesis and tumor aggressiveness, and produces more significant anti-tumor effect than single agent by blocking tumor growth and impacting the ability of the tumor to recover upon withdrawal of the therapy. [4]

Protocol

Kinase Assay:

[1]

+ Expand

Recombinant kinase assay and enzyme kinetics :

Briefly, purified-activated FAK kinase domain (amino acid 410–689) is reacted with 50 μM ATP and 10 μg per well of a random peptide polymer of Glu and Tyr, p(Glu/Tyr), in kinase buffer [50 mM HEPES (pH 7.5), 125 mM NaCl, and 48 mM MgCl2] for 15 minutes. Phosphorylation of p(Glu/Tyr) is challenged with serially diluted PF-562271 at 1/2-Log concentrations starting at a top concentration of 1 μM. Each concentration is tested in triplicate. Phosphorylation of p(Glu/Tyr) is detected with a general antiphospho-tyrosine (PY20) antibody followed by horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG antibody. HRP substrate is added, and absorbance readings at 450 nm are obtained after addition of stop solution (2 M H2SO4). IC50 values are determined using the Hill-Slope Model. Broad kinase selectivity profiling is performed in house and by using the KinaseProfiler Selectivity Screening Service available through UpState Biotechnology.
Cell Research:

[2]

+ Expand
  • Cell lines: Squamous cell carcinoma (SCC)
  • Concentrations: 0 to 1 μM
  • Incubation Time: 72 hours
  • Method:

    Cells are plated for 48 hours before addition of PF-562271. After 3 days cells are fixed by addition of ice cold 25% trichloroacetic acid (TCA) solution prior to staining with Sulforhodamine B (SRB) dye solution. Plates are washed with 1% glacial acetic acid, air-dried and resuspended in 10 mM Tris buffer, pH 10.5 before reading absorbance at 540 nm. Curve fitting and generation of IC50 values is carried out using GraphPad Prism 4 software from six replicates.


    (Only for Reference)
Animal Research:

[1]

+ Expand
  • Animal Models: PC-3M, BT474, BxPc3, LoVo, U87MG, H125 and H460 cells are injected s.c. into the right flank of athymic female mice .
  • Formulation: PF-562271 is dissolved in 5% Gelucire.
  • Dosages: ≤100 mg/kg
  • Administration: Administered via p.o.
    (Only for Reference)

Solubility (25°C)

In vitro DMSO 14 mg/mL warmed (21.03 mM)
Water Insoluble
Ethanol Insoluble
In vivo Add solvents to the product individually and in order(Data is from Selleck tests instead of citations):
4% DMSO+30% PEG 300+ddH2O
For best results, use promptly after mixing.
3mg/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 665.66
Formula

C21H20F3N7O3S.C6H6O3S

CAS No. 939791-38-5
Storage powder
in solvent
Synonyms PF-562271 Besylate

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Clinical Trial Information

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT00666926 Completed Head and Neck Neoplasm|Prostatic Neoplasm|Pancreatic Neoplasm Verastem Inc. December 2005 Phase 1

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

Tel: +1-832-582-8158 Ext:3

If you have any other enquiries, please leave a message.

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Frequently Asked Questions

  • Question 1:

    We are planning both in vitro and in vivo experiments and want to know how to reconstitute the drug for these purposes?

  • Answer:

    PF-00562271 has poor solubility in DMSO and water. Its solubility in DMSO is only 0.4mg/ml. In a previous literature report (http://www.ncbi.nlm.nih.gov/pubmed/18339875), the author used 5% Gelucire to formulate the compound. You can also consider other co-solvents such as PEG400, CMC, Tween80, and Captisol.

  • Question 2:

    Can you provide with a few common vehicles for PF-00562271, S2672 for use as oral gavage?

  • Answer:

    S2672 PF-00562271 can be dissolved in 0.5% CMC Na at 30 mg/ml as a suspension. If 4% DMSO can be used in your experiment, it will help dissolving the suspension more homogeneously.

FAK Signaling Pathway Map

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Cell Lines Assay Type Concentration Incubation Time Formulation Activity Description PMID