|Specific Ligand-Based Depletion of Human Albumin and IgG in the Presence of Denaturants|
Todd Sanderson, Hongshan Li, Brian Miller, Laura Alionte, and Lisa Bradbury
Poster data highlights the advantages of a ligand-based depletion method that functions in the presence of urea and is highly specific, disposable, cost effective, and flexible.
|A New Disposable, Specific Ligand-Based Method for Depletion of Albumin and IgG From Serum and Plasma|
Laura Alionte, Allison Miller, Cal Messing, Tao Hu, and Lisa Bradbury
Poster shows the advantages of a ligand-based depletion method that is highly specific, disposable, cost effective, and flexible. Pall’s Enchant Multi-Protein Affinity Separation Kit is a strong addition to the product choices for scientists engaged in biomarker discovery and proteomics research.
|Expression of stress response protein GRP78 is associated with the development of castration-resistant prostate cancer|
Llana Pootrakul, Anirban P. Mitra, Ram H. Datar, Shan-Rong Shi, Jie Cai, Debra Hawes, Susan G. Groshen, Amy S. Lee, Richard J. Cote
This study investigated the role of the GRP78 in prostate cancer progression and the development of castration-resistant prostate cancer, where cancer cells continue to survive despite the stress of an androgen-starved environment.
|Application of genetic programming in analysis of quantitative gene expression profiles for identification of nodal status in bladder cancer|
Anirban P. Mitra, Arpit A. Almal, Ben George, David W. Fry, Peter F. Lenehan, Vincenzo Pagliarulo, Richard J. Cote, Ram H. Datar, William P. Worzel
Nodal involvement in bladder cancer is an independent indicator of prognosis. This study employed an iterative machine learning process called genetic programming on quantitative expression values of 70 genes to classify primary urothelial carcinoma samples into those associated with or without nodal metastasis. The generated rules showed a strong predilection for ICAM1, MAP2K6 and KDR resulting in gene expression motifs that cumulatively suggested a pattern ICAM1>MAP2K6>KDR for node positive ca
|How to Perform High Throughput siRNA Transfection|
Mark Hewitson, Hanno Hermann and Neil Benn
We demonstrate the optimization of siRNA transfection process for time and costs in a high throughput application with two independent measurements of the RNAi effect: The visual monitoring of Eg5 expression and the quantitative PCR measurement of GAPDH mRNA after transfection of Hela cells. The results show, that 1 wash cycle is sufficient for the removal of any remnants of the transfection complex from disposable tips, making the tips re-usable.
|New Protein Biomarkers for Histopathological Classification of Breast Cancer |
Brozkova Kristyna, Knoflickova Dana, Bouchal Pavel, Nenutil Rudolf and Vojtesek Borivoj
We used Ciphergen technology based on SELDI TOF-MS (Surface Enhanced Laser Desorption/Ionisation Time-of-Flight Mass Spectrometry) for analysis of protein expression profiles of 105 breast cancer tissue samples. The aim of this study was the identification of single proteins or protein patterns specific for different tumour subgroups, which should take advantage of the biomarker as an alternative to commonly used diagnostic and prognostic characteristics.
|Development of an Automated siRNA Screening of Host Macrophages Genes Involved in Mycobacterium Tuberculosis Infection|
Jean-Philippe Carralot, Chang-Bok Lee, Annette Boese, Boris Lenseigne, Auguste Genovesio, Peter Sommer, Thierry Christophe and Priscille Brodin
In order to identify host genes required for M. tuberculosis infection and persistence, we developed a phenotypic cell-based assay in both murine and human cells and adapted it for high throughput and high content screening purposes. Knock-down efficiencies above 80% were achieved in “hard-to-Transfect” macrophage cells. Validation of the assay performed with control siRNAs will be discussed.
|Quantification of siRNA by a Novel Competitive-qPCR Method|
Wei-li, Liu., Mark Stevenson and Len Seymour
We have developed a competitive qPCR method in which siRNA competes with a homologous forward primer to bind template DNA, giving siRNA concentration dependent inhibition. The addition of E6-siRNA to cqPCR led to inhibition of amplification in a linear concentration-dependent manner, with as little as 200pg of siRNA capable of being detected. Irrelevant siRNA had no effect on amplification confirming specificity
|The Silencing of Multidrug Resistance-Associated Protein 5 by siRNA Complexes|
M.Malinen, E.Mannermaa, T.Ryhänen, E.Kuusela, M.Häkli, M.Yliperttula and A.Urtti
The purpose is to study the role of multidrug resistance-associated protein 5, MRP5 in drug metabolism in human retinal pigment epithelium (RPE). RPE forms the outer part of blood-retinal barrier (BRB) which restricts movements of solutes from systemic bloodstream to the neural retina. The efflux protein, MPR5 is expressed in RPE but its functions are mainly unknown. SiRNA will be tested as a tool to clarify the role of MRP5.