Journal: Proteomics

Article Title: Identification, prioritization and evaluation of glycoproteins for aggressive prostate cancer using quantitative glycoproteomics and antibody-based assays on tissue specimens

doi: 10.1002/pmic.201200541

Figure Lengend Snippet: Standard Curve of ELISA assays for protein quantification of cartilage oligomeric matrix protein (COMP), periostin, membrane primary amine oxidase (VAP-1) and cathepsin L. A), dose-response curves for COMP. B), dose-response curve for periostin. C), dose-response curve for VAP-1. D), dose-response curve for cathepsin L.

Article Snippet: Materials Hydrazide resin and sodium periodate were from Bio-Rad (Hercules, CA); sequencing-grade trypsin and TMB reagent were from Promega (Madison, WI); PNGase F was from New England Biolabs (Ipswich, MA); C18 columns were from Waters (Milford, MA); Recombinant protein, capture and detection antibody of human cartilage oligomeric matrix protein (COMP), periostin, cathepsin L, clusterin and galectin-3 binding protein, streptavidin-HRP conjugates, ELISA plates and human VAP-1 Quantikine ELISA Kit were from R&D systems (Minneapolis, MN); All other chemicals were from Sigma-Aldrich (St. Louis, MO).

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Biophysical Journal

Article Title: Detecting Protein Aggregates on Untreated Human Tissue Samples by Atomic Force Microscopy Recognition Imaging

doi: 10.1016/j.bpj.2010.06.044

Figure Lengend Snippet: AFM images of normal lens capsule mounted on a glass slide using MAC mode in PBS with injection of anti-clusterin antibody (150 μg/mL) to block surface-bound clusterin. On the left are topography images whereas the right side is the corresponding recognition images. All images are acquired at a tip oscillation amplitude of 44 nm. (A and B) Acquired before addition of antibody. (C and D) Acquired 30 min after injection. (E and F) Acquired 50 min after injection. The arrows point out corresponding topographical (left) and recognition (right) features. Scale bars = 300 nm.

Article Snippet: Sample preparation for AFM For force spectroscopy measurements, freshly cleaved mica was incubated with recombinant human clusterin (human recombinant clusterin; Biomedica Medizinprodukte, Austria; 150 μ g/mL in PBS buffer) for 1 h, and then rinsed 50 times in 500 μ L PBS buffer to remove any loose protein.

Techniques: Injection, Blocking Assay

Journal: Biophysical Journal

Article Title: Detecting Protein Aggregates on Untreated Human Tissue Samples by Atomic Force Microscopy Recognition Imaging

doi: 10.1016/j.bpj.2010.06.044

Figure Lengend Snippet: Confocal microscopy images of the anterior side of: (A) normal lens capsule (scale bar = 20 μm) with inset of digital zoom (scale bar = 500 nm); and (B) PEX lens capsule (scale bar = 20 μm) with inset of digital zoom (scale bar = 500 nm) immunolabeled with the anti-clusterin primary antibody and labeling detected with Alex Flour-488 conjugated anti-rabbit IgG secondary antibody. Image brightness curves adjusted for fluorescence visibility. Yellow arrows denote the edge of the tissue, and the cyan arrows denote the area of the image occupied by tissue. In A, this edge is due to the pinning procedure used during labeling, whereas B is the edge of the surgical cut of the lens capsule.

Article Snippet: Sample preparation for AFM For force spectroscopy measurements, freshly cleaved mica was incubated with recombinant human clusterin (human recombinant clusterin; Biomedica Medizinprodukte, Austria; 150 μ g/mL in PBS buffer) for 1 h, and then rinsed 50 times in 500 μ L PBS buffer to remove any loose protein.

Techniques: Confocal Microscopy, Immunolabeling, Labeling, Fluorescence