rabbit polyclonal anti-fpgs  (Spring Bioscience)

Journal: Oncology Reports

Article Title: High pemetrexed sensitivity of docetaxel-resistant A549 cells is mediated by TP53 status and downregulated thymidylate synthase

doi: 10.3892/or.2017.5951

Figure Lengend Snippet: Characterization of the effects of TS and GGH expression on PEM sensitivities. The expression levels of folate synthesis enzymes were examined by western blot analysis in DOC-resistant A549 sublines. The enzymes of TS and DHFR are associated with de novo synthesis, and TK1 is associated with the salvage pathway of DNA synthesis. FPGS and GGH are poly/mono-glutamylation counteracting enzymes. Anti-β-actin was used as a control for loading. Representative data are shown as follows: (A) overexpression of TS in A549/D16 was detected using (B) western blotting with PEM sensitivities determined by (C) MTT assay. Similar procedures were performed with (D) GGH overexpression and the (E) effect on PEM sensitivity. TS, thymidylate synthase; GGH, γ-glutamyl hydrolase; PEM, pemetrexed; DOC, docetaxel; DHFR, dihydrofolate reductase; FPGS, folylpolyglutamate synthase.

Article Snippet: Proteins (10–30 μg) transferred onto polyvinylidene fluoride (PVDF) membranes were reacted with polyclonal anti-TS (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), anti-DHFR, FPGS, GGH and TK1 (GeneTex, Irvine, CA, USA), anti-β-actin (NeoMarker, Fremont, CA, USA) and anti-TP53 (Dako, Carpinteria, CA, USA) separately, followed by conjugation of anti-rabbit (Santa Cruz Biotechnology, Inc.) or anti-mouse (Calbiochem, La Jolla, CA, USA) IgG to horseradish peroxidase.

Techniques: Expressing, Western Blot, DNA Synthesis, Control, Over Expression, MTT Assay