EMAIL THIS PAGE TO A FRIEND

PloS one

Reliable quantification of protein expression and cellular localization in histological sections.


PMID 25013898

Abstract

In targeted therapy, patient tumors are analyzed for aberrant activations of core cancer pathways, monitored based on biomarker expression, to ensure efficient treatment. Thus, diagnosis and therapeutic decisions are often based on the status of biomarkers determined by immunohistochemistry in combination with other clinical parameters. Standard evaluation of cancer specimen by immunohistochemistry is frequently impeded by its dependence on subjective interpretation, showing considerable intra- and inter-observer variability. To make treatment decisions more reliable, automated image analysis is an attractive possibility to reproducibly quantify biomarker expression in patient tissue samples. We tested whether image analysis could detect subtle differences in protein expression levels. Gene dosage effects generate well-graded expression patterns for most gene-products, which vary by a factor of two between wildtype and haploinsufficient cells lacking one allele. We used conditional mouse models with deletion of the transcription factors Stat5ab in the liver as well Junb deletion in a T-cell lymphoma model. We quantified the expression of total or activated STAT5AB or JUNB protein in normal (Stat5ab+/+ or JunB+/+), hemizygous (Stat5ab+/Δ or JunB+/Δ) or knockout (Stat5abΔ/Δ or JunBΔ/Δ) settings. Image analysis was able to accurately detect hemizygosity at the protein level. Moreover, nuclear signals were distinguished from cytoplasmic expression and translocation of the transcription factors from the cytoplasm to the nucleus was reliably detected and quantified using image analysis. We demonstrate that image analysis supported pathologists to score nuclear STAT5AB expression levels in immunohistologically stained human hepatocellular patient samples and decreased inter-observer variability.

Related Materials

Product #

Image

Description

Molecular Formula

Add to Cart

SAB4300311
Anti-JUNB (Ab-259) antibody produced in rabbit, affinity isolated antibody
SAB4300310
Anti-JUNB (Ab-79) antibody produced in rabbit, affinity isolated antibody
AV38135
Anti-JUNB (AB1) antibody produced in rabbit, IgG fraction of antiserum
AV100946
Anti-JUNB (AB2) antibody produced in rabbit, IgG fraction of antiserum
AV100947
Anti-JUNB (AB3) antibody produced in rabbit, IgG fraction of antiserum
AV38136
Anti-JUNB (AB4) antibody produced in rabbit, affinity isolated antibody
SAB2501262
Anti-JUNB antibody produced in goat, affinity isolated antibody, buffered aqueous solution
SAB1406056
Anti-JUNB antibody produced in mouse, purified immunoglobulin, buffered aqueous solution
HPA019149
Anti-JUNB antibody produced in rabbit, Prestige Antibodies® Powered by Atlas Antibodies, affinity isolated antibody, buffered aqueous glycerol solution
SAB4501614
Anti-JUNB antibody produced in rabbit, affinity isolated antibody
SAB1407007
Anti-RNASEH2A antibody produced in mouse, purified immunoglobulin, buffered aqueous solution
AV40646
Anti-RNASEH2A antibody produced in rabbit, affinity isolated antibody
AV40647
Anti-RNASEH2A antibody produced in rabbit, IgG fraction of antiserum
SAB1410102
Anti-RNASEH2A antibody produced in rabbit, purified immunoglobulin, buffered aqueous solution
WH0010535M1
Monoclonal Anti-RNASEH2A antibody produced in mouse, clone 3G5-F5, purified immunoglobulin, buffered aqueous solution
S4776 Somatotropin from human pituitary, recombinant, expressed in E. coli, ≥95% (SDS-PAGE), lyophilized powder