Recombinant Anti-ERG antibody [EPR3864] (ab92513)

Formalin-fixed, paraffin-embedded mouse brain tissue stained for ERG using ab92513 at 1/200 dilution in immunohistochemical analysis. A horse radish peroxidase antibody was used as the secondary antibody.

Antigen Retrieval: 40x; Proteinase K antigen retrieval - 15 min at 37 C

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

ab92513 staining ERG in THP-1 cells, with negative expression in HCT116 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% Triton x-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab92513 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 μg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

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

Intracellular 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 Fluorr^® 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.

Alexa Fluor^® 488 (ab196374) and Alexa Fluor^® 647 (ab196149) conjugated versions are available for this clone.

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

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.