Category: Discussion Topics

Construction and Analysis of Glycan Microarrays

Protein glycosylation is a complex post-translational modification (PTM) that provides great proteomic diversity. It involves the attachment of sugars at specific amino acid residues. Asn (N-linked) or Ser/Thr (O-linked) being the residues most frequently glycosylated. Glycosylation is critical for physiological and pathological cellular functions and affects the bioavailability, efficacy, stability, and solubility of therapeutic glycoproteins. … Continued

Flow chambers with precise control of environmental conditions for single-cell assays

A simple fluidic design integrates directly into standard cell culture and biochemical assays as a part of new experimental methods published recently. The fluidic design used single channel or T-shaped flow chambers and a syringe pump to effectively culture and assay cells in a continuous gradient of conditions. These methods simultaneously expand experimental capability and … Continued

The Trafficking of wheat prolamins in rice endosperm

Wheat is the most important crop in Europe due in part to high relative crop yields over a range of environments. “(Wheats) success as a food crop is also due to the unique properties which form the gluten fraction… It (is) necessary to understand the pathways and mechanisms which determine the synthesis, processing, trafficking and … Continued

Antibody Validation Assay with PATH® Protein Microarray Substrate

The analysis of antibody interactions to 19,500 recombinant full-length human proteins featured on HuProt™ Human Proteome Microarray provides a robust and rapid method to evaluate antibodies’ quality. Introduction Antibodies are the most commonly used research tools. Applied in basic research as well as in clinical and diagnostic medicine, their quality, consistency, and availability are fundamental … Continued

Confocal Fluorescence Microscopy of CRISPR-generated G6PD Knockout mouse liver cells (TIB-73)

CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) Cas-9 knockout of glucose 6-phosphate dehydrogenase (G6PD) The first surviving G6PD- knockout somatic cell line was generated with CRISPR-Cas-9 technology to support subcellular localization studies. G6PD,  glucose 6-phosphate dehydrogenase, is the rate-limiting enzyme in the pentose phosphate pathway. G6PD is involved in oxidative stress, apoptosis, angiogenesis and cancer … Continued

Epitope Mapping Using Peptide Microarray

Epitope mapping allows the identification of polyclonal antibodies produced as a response to cytotoxins, allergens, neuronal or inflammatory conditions and, in autoimmune diseases to autoantigens. Routine assays for autoimmune response profiling are generally performed by ELISAs and fluorescence immunoassays. Individual assays are performed in microtiter plates, with each well representing a single antigen. Peptide microarrays provide … Continued

Applications of Protein Microarrays for Biomarker Discovery.

In a recent paper published in the Journal of Neuroinflammation the immune response associated with elevated intermittent ocular hypertension has been evaluated.  An antigen microarray assay was performed with ten different antigens spotted on ONCYTE®  nitrocellulose film slides and used to probe serum from an intermittent ocular hypertension animal model. High Intraocular pressure is a main risk … Continued

Reverse Phase Protein Array Used to Elucidate Pro-inflammatory Signalomes Provides an Excellent Guide for RPPA Methodologies and Techniques

Application of Reverse-Phase Protein Arrays (RPPA) has primarily centered on the discovery and identification of cell signaling pathways involved in cancer development and progression. The RPPA approach and technology has found expanded utility in other disciplines such as in the study of inflammatory diseases.Recently Negm, et al. employed RPPA to study the signaling pathways downstream … Continued

Routine Clinical Adoption of Immunoassay Multiplexing, Whether Microarray or Cytometric Bead Array, Is Still Several Years Away Due to Operational Entrenchment

An objective look at immunoassay multiplexing, where it stands today and contrasted to the standard ELISA was authored recently by Dr. Lucy Fairclough et al. The discussion argued the efficacy of multiplexing as well as the pros and cons of present methods available.Commercial availability of multiplex immunoassays for life science discovery research is rapidly expanding … Continued

Review of Mujawar et al Analytical Chemistry – Distribution of Biomolecules in Porous Nitrocellulose Films

An interesting paper in (Analytical Chemistry) describes the use of confocal fluorescence section imaging to analyze protein distribution in microarray spots on nitrocellulose film slides. By scanning and imaging through the Z dimension of a printed spot of fluor labeled protein, a 3-dimensional quantitation of fluorophore density in the films was obtained, showing that commercial … Continued