Journal: Oncotarget

Article Title: PRDX2 and PRDX4 are negative regulators of hypoxia-inducible factors under conditions of prolonged hypoxia

doi: 10.18632/oncotarget.7142

Figure Lengend Snippet: A. and B. HeLa cells were transfected with PRDX2-V5 (A) or PRDX4-V5 (B) vector and WCL was incubated with purified GST or GST-HIF-1α fusion protein in the presence of glutathione-Sepharose beads, followed by immunoblot assays with anti-V5 antibody (upper panels) or Ponceau S staining (lower panels).

Article Snippet: Whole cell lysates were prepared in modified RIPA buffer [50 mM Tris-HCl (pH 7.5), 1 mM β-mercaptoethanol, 150 mM NaCl, 1% Igepal, and protease inhibitor cocktail] and incubated overnight with the following antibodies (catalog number and supplier): HIF-1α (sc-10790, Santa Cruz); V5 epitope (NB600-381, Novus Biologicals), HIF-2α (NB100-122, Novus Biologicals), or p300 (NB500-161, Novus Biologicals) in the presence of protein A Sepharose beads (NBP1-97240, Novus Biologicals).

Techniques: Transfection, Plasmid Preparation, Incubation, Purification, Western Blot, Staining

Journal: Translational Vision Science & Technology

Article Title: Detection of TTR Amyloid in the Conjunctiva Using a Novel Fluorescent Ocular Tracer

doi: 10.1167/tvst.13.2.11

Figure Lengend Snippet: Spectroscopic and binding properties of AMDX-9101 combined with aggregated wild type TTR. ( A ) Fluorescent emission of AMDX-9101 before ( blue ) and after ( purple ) mixing with TTR aggregates. ( B ) Plot of the fluorescence intensity (λ max = 545 nm) as a function of the concentration of AMDX-9101 in the presence of aggregated TTR peptides (5 µM) in solution. Fitting this data to the equation: y = Bx/(Kd + x) revealed a Kd of 1.476 ± 0.3452 µM for association of AMDX-9101 to aggregated TTR peptides. ( C ) Spectroscopic properties of AMDX-9101 and wt TTR. Fold increase determined from the emission with 4 µM AMDX and 5 µM aggregated TTR at the maximum emission of AMDX + TTR.

Article Snippet: TTR aggregates were prepared as described by Frangolho et al. Lyophilized TTR (Novus Biologicals NBP2-62523) was resuspended in 50 mM glycine buffer as a 100 μM stock.

Techniques: Binding Assay, Fluorescence, Concentration Assay