Recombinant Anti-ERG antibody [EPR3864] (ab92513)

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST

Functional characterization and detection of genetic alterations in GEDI-captured cells. The TMPRSS2:ERG fusion protein is detected in GEDI-captured circulating tumor cells (CTCs) from a castrate-resistant prostate cancer (CRPC) patient. PSMA-captured CTCs were stained on the device with ab92513. Representative examples of PSMA+/CD45− CTCs are shown, two of which are positive for ERG. Scale bars: 10 microns.

ERG and GSTP1 immunostainings of human prostate cancer samples using ab92513.

Representative immunohistochemical images of prostate cancer samples are shown that were positive for ERG and negative for GSTP1 (A), positive for both ERG and GSTP1 (B), negative for both ERG and GSTP1(C), and negative for ERG and positive for GSTP1 (D). The internal staining control for ERG is the endothelium (arrows) and for GSTP1 the stromal and/or basal cells of normal prostate glands. N, normal prostate gland; S, Stroma; T, tumor gland. Scale bars equal 100μm

Flow Cytometry analysis of THP-1 (human monocytic leukemia cell line) cells labeling ERG with purified ab92513 at 1:1000 dilution(1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488)(ab150077)(1:2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST

Immunofluorescence staining of THP-1 (human monocytic leukemia cell line) cells with purified ab92513 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab92513 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

ab92513 staining ERG in Human brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 3% serum for 30 minutes at 25°C. Samples were incubated with primary antibody (1/100 in 3% Horse serum) for 15 hours at 4°C. An undiluted HRP-conjugated Horse anti-rabbit IgG polyclonal was used as the secondary antibody.

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Immunohistochemical analysis of Fus A5 transgenic mouse prostate tissue, staining ERG with unpurified ab92513.

Tissue sections were blocked in 20% goat serum, 2% BSA in PBS (GSB) for 40 min at room temperature, followed by an overnight incubation in primary antibody (1/25 in GSB) at 4°C. Following washes, the sections were incubated with a fluorescently tagged secondary antibody for 30 min at room temperature.

Immunohistochemical analysis of paraffin embedded Human Prostatic adenocarcinoma stage 3 tissue using unpurified ab92513 showing +ve staining.