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Ab10425

Manufactured by Abcam
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Ab10425 is a primary antibody product. It is a monoclonal antibody that recognizes a specific target protein. The antibody is designed for research use in standard immunochemical applications.

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Lab products found in correlation

Ripa buffer, by Thermo Fisher Scientific (2 mentions) Anti phospho frs2 α tyr196, by Cell Signaling Technology (2 mentions) Anti gapdh, by Cell Signaling Technology (1 mentions)

3 protocols using ab10425

1

Characterizing FGFR1c Phosphorylation of FRS2

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We first evaluated the effect of the V429E mutation on the ability of FGFR1c to phosphorylate FRS2 using cell-based assay. WT and V429E FGFR1c were cloned into the lentiviral vector FUCRW following standard protocols. BaF3 cells were maintained as described23 (link) and transfected with FGFR1 pseudoviral stock in Hank's Balanced Salt Solution buffer. Stably transduced cells were treated with 1.5 nM of FGF1 for 10 min, rinsed in PBS and then lysed in RIPA buffer (Thermo Scientific). Cell extract (30 μg) was resolved by SDS-PAGE and analyzed by western blotting using anti FGFR1 (inhouse antibody raised in rabbit), anti-FRS2 (Abcam ab10425) and anti Phospho-FRS2-α (Tyr196) (Cell Signaling, #3864) antibodies.
Villanueva C., Jacobson-Dickman E., Xu C., Manouvrier S., Dwyer A.A., Sykiotis G.P., Beenken A., Liu Y., Tommiska J., Hu Y., Tiosano D., Gerard M., Leger J., Drouin-Garraud V., Lefebvre H., Polak M., Carel J.C., Phan-Hug F., Hauschild M., Plummer L., Rey J.P., Raivio T., Bouloux P., Sidis Y., Mohammadi M., de Roux N, & Pitteloud N. (2014). Congenital hypogonadotropic hypogonadism with split hand/foot malformation: a clinical entity with a high frequency of FGFR1 mutations. Genetics in medicine : official journal of the American College of Medical Genetics, 17(8), 651-659.
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2

Western Blot Analysis of FGF Signaling

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Western blot analysis was conducted using 20 μg of cell lysates in radioimmunoprecipitation assay buffer and the proteins were examined with monoclonal antibodies directed against FGF19 (83 348), p‐FRS2 (3961), AKT (11 962), p‐AKT (9271), ERK (4695), p‐ERK (12 638), cleaved‐Caspase 3 (CC3; 9661) and anti‐GAPDH (5174, Cell Signaling Technology, Danvers, MA, USA), and FRS2 (ab10425), FGFR4 (ab5841), and p‐FGFR4 (ab192589, Abcam, Cambridge, UK).
Zhang X., Kong M., Zhang Z., Xu S., Yan F., Wei L, & Zhou J. (2017). FGF19 genetic amplification as a potential therapeutic target in lung squamous cell carcinomas. Thoracic Cancer, 8(6), 655-665.
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3

Characterizing FGFR1c Phosphorylation of FRS2

Check if the same lab product or an alternative is used in the 5 most similar protocols
We first evaluated the effect of the V429E mutation on the ability of FGFR1c to phosphorylate FRS2 using cell-based assay. WT and V429E FGFR1c were cloned into the lentiviral vector FUCRW following standard protocols. BaF3 cells were maintained as described23 (link) and transfected with FGFR1 pseudoviral stock in Hank's Balanced Salt Solution buffer. Stably transduced cells were treated with 1.5 nM of FGF1 for 10 min, rinsed in PBS and then lysed in RIPA buffer (Thermo Scientific). Cell extract (30 μg) was resolved by SDS-PAGE and analyzed by western blotting using anti FGFR1 (inhouse antibody raised in rabbit), anti-FRS2 (Abcam ab10425) and anti Phospho-FRS2-α (Tyr196) (Cell Signaling, #3864) antibodies.
Villanueva C., Jacobson-Dickman E., Xu C., Manouvrier S., Dwyer A.A., Sykiotis G.P., Beenken A., Liu Y., Tommiska J., Hu Y., Tiosano D., Gerard M., Leger J., Drouin-Garraud V., Lefebvre H., Polak M., Carel J.C., Phan-Hug F., Hauschild M., Plummer L., Rey J.P., Raivio T., Bouloux P., Sidis Y., Mohammadi M., de Roux N, & Pitteloud N. (2014). Congenital hypogonadotropic hypogonadism with split hand/foot malformation: a clinical entity with a high frequency of FGFR1 mutations. Genetics in medicine : official journal of the American College of Medical Genetics, 17(8), 651-659.
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