Immunohistochemical Evaluation of ERG Expression

TMA tissue sections (2.5 μm) were deparaffinized and epitopes were retrieved using TEG buffer, as previously described [60 (link)]. TMA sections were stained with rabbit monoclonal ERG antibody (2805–1, Epitomics) in a 1:150 dilution in TEG. Secondary staining was performed using the EnVision + System (HRP Labelled Polymer Anti-Rabbit K4003, DakoCytomation). For all cores, ERG immunoreactivity (0,no staining; 1, moderate; 2, strong) was evaluated and scored by two independent observers (SH and ASL) as described above. Positive staining was used as a proxy for ERG fusion status as previously described [45 (link), 46 (link)].

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Strand S.H., Hoyer S., Lynnerup A.S., Haldrup C., Storebjerg T.M., Borre M., Orntoft T.F, & Sorensen K.D. (2015). High levels of 5-hydroxymethylcytosine (5hmC) is an adverse predictor of biochemical recurrence after prostatectomy in ERG-negative prostate cancer. Clinical Epigenetics, 7, 111.

Publication 2015

EnVision + System (HRP Labelled Polymer Anti-Rabbit K4003,

Buffer Dilution Epitopes Monoclonal antibody Polymer Rabbit Tissue

Corresponding Organization :

Other organizations : Aarhus University Hospital

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Variable analysis

independent variables

Epitope retrieval using TEG buffer
Antibody concentration (1:150 dilution of rabbit monoclonal ERG antibody)

dependent variables

ERG immunoreactivity (scored as 0, 1, or 2)

control variables

TMA tissue sections (2.5 μm)
Deparaffinization of TMA sections
Secondary staining using EnVision + System (HRP Labelled Polymer Anti-Rabbit K4003)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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