A 419259 trihydrochloride, Inhibitor | A419259, inhibitor

A 419259 trihydrochloride

Cat. Num. AAA5764388

• Purity: 98.00%
• Synonyms: A 419259 trihydrochloride; A419259; inhibitor

• Purity/Purification: 98.00%
• Form/Format: Solid. Powder

• Solubility: Water: 53mg/mL (89.53mM); (<1mg/ml refers to the product slightly soluble or insoluble)
• Formula: C29H37Cl3N6O
• SMILES: CN1CCN([C@@]2([H])CC[C@@](N3C=C(C(C3=NC=N4)=C4N)C5=CC=C(OC6=CC=CC=C6)C=C5)([H])CC2)CC1.Cl.Cl.Cl
• CAS Number: 1435934-25-0
• Cell Research: K-562 cells are grown in RPMI 1640 supplemented with 10% fetal calf serum (FCS), and 50 g/mL Gentamycin. Meg-01 cells are cultured in Vitacell modified RPMI 1640 (ATCC), supplemented with 10% FCS and 50ug/mL Gentamycin. The human GM-CSF-dependent myeloid leukemia cell line TF-1 is grown in RPMI 1640 supplemented with 10% FCS, 50ug/mL Gentamycin, and 1ng/mL of recombinant human GM-CSF. DAGM murine myeloid leukemia cells are cultured in RPMI 1640 supplemented with 10% FCS, 50ug/mL Gentamycin, and 0.5ng/mL recombinant IL-3. Concentrated stock solutions of PP2 (5mM) and A-419259 (2mM) are prepared in DMSO and stored at -20 degree C. Cellular proliferation is measured using the Live/Dead growth assay. This assay employs calcein-AM, a fluorogenic esterase substrate that is taken up by viable cells and hydrolyzed intracellularly, releasing a green fluorescent product. Briefly, 10^4 cells are plated per well in 96-well plates for each day of a 4-day time course. Various concentrations of PP2, A-419259 or vehicle control are added to the wells (five wells per concentration per day) and the plates are incubated at 37 degree C. At each time point, one plate is centrifuged at 1500g for 10 min to pellet the cells. Cells are washed with PBS, and calcein-AM is added to each well to a final concentration of 1uM. Plates are incubated in the dark at room temperature for 1 hours. The plates are then read at 485/530nM (excitation/emission) using a fluorescent plate reader and data are analyzed with SoftMax Pro software [1].
• Kinase Assay: In vitro kinase assays are performed on His(6)-tagged Lck (residues 62-509) and full-length c-Abl purified from Sf-9 cells, and commercial sources of Lyn, Src and PKC. Lck, Lyn, Src and Abl activities are measured with an ELISA-based assay. Flat bottom 96-well ELISA plates are incubated with a 200ug/mL solution of Poly(Glu, Tyr) 4:1 substrate in phosphate-buffered saline (PBS) for 1 hour at 37 degree C and then washed with PBS containing 0.1% Tween-20 (PBS-T). Inhibitor dilutions are added to the washed plates already containing the appropriate enzyme in kinase assay buffer (250mM Mopso, pH 6.75, 10mM MgCl2, 2mM MnCl2, 2.5mM DTT, 0.02% BSA, 2mM Na3VO4, 5% DMSO, 5uM ATP). After incubation at room temperature for 20 min, plates are washed three times with PBS-T and plate-bound phosphotyrosine is detected with an anti-phosphotyrosine-HRP antibody conjugate and subsequent color development with K-Blue reagents [1].
• Preparation and Storage: Store at -20 degree C for 3 years powder; -80 degree C for 2 years in solvent

Related Product Information for A419259, inhibitor

A 419259 trihydrochloride is an Src family kinases inhibitor (IC50s: 9nM, 3nM and 3nM for Src, Lck and Lyn).

Product Categories/Family for A419259, inhibitor

Angiogenesis; Src; Tyrosine Kinase/Adaptors

References

Wilson MB, et al. Selective pyrrolo-pyrimidine inhibitors reveal a necessary role for Src family kinases in Bcr-Abl signal transduction and oncogenesis. Oncogene. 2002 Nov 21;21(53):8075-88.
Wilson MB, et al. Selective pyrrolo-pyrimidine inhibitors reveal a necessary role for Src family kinases in Bcr-Abl signal transduction and oncogenesis. Oncogene. 2002 Nov 21;21(53):8075-88.
Pene-Dumitrescu T, et al. An inhibitor-resistant mutant of Hck protects CML cells against the antiproliferative and apoptotic effects of the broad-spectrum Src family kinase inhibitor A-419259.Oncogene. 2008 Nov 27;27(56):7055-69.
Pene-Dumitrescu T, et al. An inhibitor-resistant mutant of Hck protects CML cells against the antiproliferative and apoptotic effects of the broad-spectrum Src family kinase inhibitor A-419259.Oncogene. 2008 Nov 27;27(56):7055-69.