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Effective Screening of Inhibitors for Influence on Proliferation and Survival of Mouse Neural Stem Cells.

Rapid Screening of Receptor Tyrosine Kinase (RTK) Inhibition Using InhibitorSelect™ Inhibitor Library I and WideScreen™ RTK pTyr and Total 7-plex Assay Kits on a Luminex® xMAP® Platform.

InhibitorSelect 96-Well Protein Kinase Inhibitor Libiraries I & II were screened for influence on the proliferation and survival of the mouse neural stem cells (mNS) in an ATP bioluminescence cell viability assay. mNS cells were plated at 3,000 cells/well and viability assay performed after 2 days of incubation. Each compound was tested in each of four growth factor backgrounds: (A) No GFs – No Growth Factors (to identify survival/ proliferation factors), (B) Sub EGF – Sub-optimal EGF (to identify inhibitors/ potentiators) 20 pg/mL EGF, (C) Sub FGF2 – Suboptimal FGF2 (to identify inhibitors/ potentiators) 500 pg/mL FGF2, (D) Max GFs – Maximal EGF + FGF2 (to identify inhibitors/ potentiators) 20 ng/mL EGF + 20 ng/mL FGF2. Results for 20 inhibitors are presented as mean data (of n-4 wells per condition) with error bars indicating standard error of the mean (SEM). The presence of inhibitor K-252a, Nocardiospsis sp. (Cat. No. 420297) alone in the culture media resulted in a 10-fold mNS cell viability. Data courtesy of Donna McLaren, Stem Cell Sciences, Cambridge, UK.
Click image to enlarge.
InhibitorSelect 96-Well Protein Kinase Inhibitor Libiraries I & II (160 inhibitors; Cat. No. 539744 and 539745) were screened for influence on the proliferation and survival of the mouse neural stem cells (mNS) in an ATP bioluminescence cell viability assay. mNS cells were plated at 3,000 cells/well and viability assay performed after 2 days of incubation. Each compound was tested in each of four growth factor backgrounds:
  • (A) No GFs – No Growth Factors (to identify survival/ proliferation factors)
  • (B) Sub EGF – Sub-optimal EGF (to identify inhibitors/ potentiators) 20 pg/mL EGF
  • (C) Sub FGF2 – Suboptimal FGF2 (to identify inhibitors/ potentiators) 500 pg/mL FGF2
  • (D) Max GFs – Maximal EGF + FGF2 (to identify inhibitors/ potentiators) 20 ng/mL EGF + 20 ng/mL FGF2
Results for 20 inhibitors are presented as mean data (of n-4 wells per condition) with error bars indicating standard error of the mean (SEM). The presence of inhibitor K-252a, Nocardiospsis sp. (Cat. No. 420297) alone in the culture media resulted in a 10-fold mNS cell viability.

Data courtesy of Donna McLaren, Stem Cell Sciences, Cambridge, UK

Over twenty kinase inhibitors were rapidly screened for their ability to inhibit RTK activation using the InhibitorSelect 96-well Inhibitor Library I. Cell lines were stimulated with the select growth factors or mitogens. Following stimulation, the activation and inhibition of several TRKs (EGFR, HER2, HGFR, IGF1R, PDGFR, TIE2, and VEGFR2), were measured using the WideScreen RTK pTyr 7-plex Assay. RTK phosphorylation levels were normalized against total RTK concentrations using WideScreen™ RTK Total 7-plex Assay. Representative data for VEGFR2 are shown.
Click image to enlarge.
Over twenty kinase inhibitors were rapidly screened for their ability to inhibit RTK activation using the InhibitorSelect 96-well Inhibitor Library I (Cat. No. 539744). Cell lines were stimulated with the select growth factors or mitogens. Following stimulation, the activation and inhibition of several TRKs (EGFR, HER2, HGFR, IGF1R, PDGFR, TIE2, and VEGFR2), were measured using the WideScreen RTK pTyr 7-plex Assay (Cat. No. 71943). RTK phosphorylation levels were normalized against total RTK concentrations using WideScreen RTK Total 7-plex Assay (Cat. No. 71942). Representative data for VEGFR2 are shown.

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Characteristics of three Akt inhibitors included in InhibitorSelect Libraries are shown to demonstrate the diversity and quality of inhibitors provided.

Akt Inhibitor V, Triciribine (Cat. No. 124012)


Akt Inhibitor V, Triciribine


Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 (Cat. No. 124018)


Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2


Akt Inhibitor X (Cat. No. 124020)


Akt Inhibitor X


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