Human Phospho-FGFR1-4 (Y653/Y654) Antibody

Detection of Human Phospho- FGF R1‑4 (Y653/Y654) by Western Blot.

Western blot shows lysates of KATO-III human gastric carcinoma cell line untreated (-) or treated (+) with 100 ng/mL Recom-binant Human FGF acidic (Catalog # 232-FA) for 15 minutes. PVDF membrane was probed with 0.5 µg/mL of Rabbit Anti-Human Phospho-FGF R1-4 (Y653/Y654) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3285), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-FGF R1-4 (Y653/Y654) at approximately 120 to 145 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-FGF R1-4 (Y653/Y654) in A431 Human Cell Line.

FGF R1-4 phosphorylated at Y653/Y654 was detected in immersion fixed A431 human epithelial carcinoma cell line untreated (lower panel) or treated (upper panel) with pervanadate using Rabbit Anti-Human Phospho-FGF R1-4 (Y653/Y654) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3285) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Phospho-FGFR1-4 (Y653/Y654) by Western Blot

GNA inhibits cell proliferation in both NSCLC and erlotinib-resistant NSCLC by targeting FGFR and the downstream signaling pathways.a Cell viability was assessed using the CellTiter-Glo assay. After a 72 h treatment by GNA, CellTiter-Glo reagent was added directly to each well for a 10-min incubation. The plate was then read on a FlexStation 3 microplate luminometer. A dose−response curve was plotted, and the IC50 was calculated using GraphPad Prism software. Cell viability was assessed using MTT assay. After a 72 h treatment by GNA, 10 μl of 5 mg/ml 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) solution was added to each well and incubated for 4 h at 37 °C, then the culture medium was removed and 100 μl of DMSO was added to each well. The absorbance was measured at 570 nm on a microplate reader. A dose−response curve was plotted, and the IC50 was calculated using GraphPad Prism software. b Caspase-3/7 activity induction was evaluated after 6 h of treatment with GNA at 10, 3.33 and 1.11 μM using the Caspase-Glo 3/7 kit from Promega in the HCC827, H1650, and HCC827ER cells, respectively. The bars in the graphs represent the mean fold induction relative to the DMSO control. c H1650 cells, HCC827 cells, and HCC827ER cells were treated with GNA at various dosages for 4 h and were then probed with specified antibodies. FGFR and the downstream proteins were analyzed by western blot. d The histograms of quantitative analysis about FGFR and the downstream proteins were analyzed by western blot Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/29449529), licensed under a CC-BY license. Not internally tested by R&D Systems.