Antibody-based Proteomics for Human Tissue Profiling

Abstract

Here, we describe the use of antibody-based proteomics involving the generation of protein-specific antibodies to functionally explore the human proteome. The antibodies can be used for analysis of corresponding proteins in a wide range of assay platforms, including i) immunohistochemistry for detailed tissue profiling, ii) specific affinity reagents for various functional protein assays, and iii) capture ("pull-down") reagents for purification of specific proteins and their associated complexes for structural and biochemical analyses. In this review, the use of antibodies for such analysis will be discussed with focus on the possibility to create a descriptive and comprehensive protein atlas for tissue distribution and subcellular localization of human proteins in both normal and disease tissues.

Mitochondria: A mirror into cellular dysfunction in heart disease

“…Alternative and complementary to IMS is the use of antibody-based proteomics, which, facilitates profiling of tissue sections through antibody affinity, provides similar spatial information, but with more focus on intact proteins [230]. Unlike IMS where identification is based on the tissue components, antibody-based proteomics requires monospecific antibodies (msAb) to ensure specific interaction with the protein of interest alone.…”

“…Such an approach has been utilized for the validation and spatial localization of ovarian cancer biomarkers, showing differentiation between normal, borderline and malignant tissue [231]. The atlas proposed by Uhlen et al [230] covers a wide range of msAb probed towards a variety of tissue samples, showing the applicability of antibody-based proteomics to spatial investigations of protein expression profiles. Whilst some mitochondrial proteins have been investigated using this approach, including the 24-kDa subunit of complex 1, the impact of CV disease on these proteins has not been undertaken.…”

Sox5 can suppress platelet-derived growth factor B-induced glioma development in Ink4a-deficient mice through induction of acute cellular senescence

“…Immunohistochemical staining for SOX5 (Abcam, Cambridge, UK) of tissue microarrays including normal and cancer tissue was performed in collabration with the Swedish Human Protemome Resource Project (Uhlen and Ponten, 2005). Two samples from human brain cortex and 11 gliomas were representative and used for annotation of SOX5.…”

Selected Reaction Monitoring: A Valid Tool for Targeted Quantitation of Protein Biomarker Discovery

“…This information is either present in the repositories data base available online or have to be determined before SRM-MS experiment through bottom up shotgun LC-MS/MS experiment. Several, sites are available freely to select the target proteins (Table 1a) [8][9][10][11][12][13][14]. The success of shotgun proteomics relies on the sample preparation.…”