Anti-Human LTK Recombinant Antibody (FAM150A) (CAT#: TAB-0711CL)

Figure 1 Induction of LTK phosphorylation by FAM150A.

Figure 2 Dose-dependent induction of LTK phosphorylation by FAM150A.

Figure 3 Induction of LTK phosphorylation in the presence of FAM150A, and inhibition of FAM150A-induced LTK phosphorylation by kinase inhibitor crizotinib.

Figure 4 mRNA expression of FAM150A in various cell lines.

Figure 5 mRNA expression of FAM150A in a panel of human immune cells.

Figure 6 FAM150A and FAM150B activate ALK.

Neurite outgrowth in PC12 cells expressing either vector control, FAM150A, FAM150B, ALK, FAM150A and ALK, FAM150B and ALK or ALK-F1174L quantified in right picture. Experiments were performed in triplicate and each sample within an experiment was performed in duplicate (error bars indicate SD).

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 7 FAM150A and FAM150B activate ALK.

Whole cell lysates from PC12 cells expressing either vector control, FAM150A, ALK, FAM150A and ALK or ALK-F1174L were analyzed by immunoblot analysis of ALK, pALK-Y1604 (arrowheads), FAM150A and ERK1/2. Pan-ERK was employed for equal loading.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 8 Conditioned medium containing either FAM150A or FAM150B activates endogenous ALK.

Neurite outgrowth in PC12 cells expressing either vector control or anaplastic lymphoma kinase (ALK) were cultured in medium from Human Embryonic Kidney (HEK) 293 cells transfected with either FAM150A.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 9 Conditioned medium containing either FAM150A or FAM150B activates endogenous ALK.

Whole cell lysates from PC12 cells expressing either vector control or ALK stimulated with medium from HEK293 cells transfected with vector control, FAM150A were analyzed by immunoblot.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 10 Conditioned medium containing either FAM150A or FAM150B activates endogenous ALK.

IMR32 cells harboring a wild-type ALK receptor were stimulated for 20 min with medium from HEK293 cells transfected with either vector control, FAM150A or FAM150B prior to analysis by immunoblot.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 11 Conditioned medium containing either FAM150A or FAM150B activates endogenous ALK.

IMR32 cells harboring a wild-type ALK receptor stimulated with increased amounts of recombinant His-tagged FAM150A purified from Sf21 cells.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 12 Conditioned medium containing either FAM150A or FAM150B activates endogenous ALK.

Time course of IMR32 cells stimulated with FAM150A conditioned medium. Stimulation with ALK activating antibody mAb46 was employed as positive control.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 13 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

Binding kinetics of purified FAM150A to extracellular domain of anaplastic lymphoma kinase (ALK-ECD-Fc) in a Biacore surface plasmon resonance (SPR) analysis.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 14 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

FAM150A immunoprecipitates with human ALK. Immunoprecipitation with either anti-FLAG(DYKDDDDK)(ALK) or anti-HA (FAM150A) was performed and the resulting immunoprecipitates immunoblotted for the presence of ALK (blue arrowheads) and FAM150A (red arrowheads), *indicates immunoglobulin light and heavy chains.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 15 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

Human Embryonic Kidney (HEK) 293 cells expressing ALK were incubated with either control or HA-tagged FAM150A or HA-tagged FAM150B conditioned medium prior to analysis by immunohistochemistry. Both HA-tagged FAM150A and HA-tagged FAM150B bind to ALK-expressing cells. Higher magnification panels indicate intracellular vesicles positive for both ALK and HAtagged FAM150A/B.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 16 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

NB1 and IMR32 neuroblastoma cells were treated with 2 µg/ml monoclonal antibodies (mAB13, mAb48 or mAb135) prior to stimulation with FAM150A. Detection of ALK activation was visualized with pALK-Y1604 (arrowheads) and pERK1/2 in whole cell lysates. Pan-ERK or tubulin were employed for equal loading.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 17 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

Whole cell lysates from PC12 cells expressing either vector control or ALK-R1275Q were stimulated with medium from HEK293 cells transfected with vector control, FAM150A or FAM150B prior to analysis by immunoblot. Analysis was carried out in the presence or absence of 250 nM crizotinib. Detection of ALK activation was visualized with pALK-Y1604 (arrowheads) and pERK1/2 in whole cell lysates. Pan-ERK was employed for equal loading.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.

Figure 18 FAM150A and FAM150B bind to ALK and further activate signaling mediated by the R1275Q ALK neuroblastoma mutation.

CLB-GAR cells harboring the ALK-R1275Q mutant were stimulated for 30 min with medium from HEK293 cells transfected with either vector control, FAM150A or FAM150B prior to analysis by immunoblot. Analysis was carried out in the presence or absence of 250 nM crizotinib. Detection of ALK signaling activation was visualized with pERK1/2. Pan-ERK was employed for equal loading.

Guan, J., Umapathy, G., Yamazaki, Y., Wolfstetter, G., Mendoza, P., Pfeifer, K., ... & Halenbeck, R. (2015). FAM150A and FAM150B are activating ligands for anaplastic lymphoma kinase. elife, 4, e09811.