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

All lanes : purified

Lane 1 : rat brain lysate
Lane 2 : rat heart lysate
Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

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.

Immunohistochemical staining of paraffin embedded human kidney with purified ab92513 at a working dilution of 1/1000. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

All lanes : Anti-ERG antibody [EPR3864] (ab92513) at 1/2000 dilution (purified)

Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 55 kDa
Observed band size: 55 kDa

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.

Anti-ERG antibody [EPR3864] (ab92513) at 1/1000 dilution (unpurified) + Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate at 10 µg

Secondary
HRP labelled Goat anti-Rabbit at 1/2000 dilution

Predicted band size: 55 kDa

Lanes 1 & 3 : Anti-ERG antibody [EPR3864] (ab92513) at 1/250 dilution (unpurified)
Lanes 2 & 4 : Anti-ERG antibody [EPR3864] (ab92513) at 1/1000 dilution (unpurified)

Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) Whole Cell Lysate
Lanes 2-4 : Jurkat Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 55 kDa
Observed band size: 55 kDa


Exposure time: 12 minutes

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.

Equilibrium disassociation constant (K_D)
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