All lanes : Anti-WDR77 antibody [EPR10708(B)] (ab154190) at 1/1000 dilution

Lane 1 : HeLa cell lysate
Lane 2 : LnCAP cell lysate
Lane 3 : 293T cell lysate
Lane 4 : fetal heart lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 37 kDa

Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labeling WDR77 with Purified ab154190 at 1/500 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

Overlay histogram showing HeLa cells stained with ab154190 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab154190, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x10⁶ cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

ab154190 staining WRD77 (green) in U2OS cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.2% Triton X-100 in PBS. Samples were incubated with primary antibody (1/150 in PBS + 5% BSA) for 1 hour at 20°C. An Alexa Fluor®488-conjugated Goat anti-rabbit IgG polyclonal (1/800) was used as the secondary antibody. Red - DAPI.

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Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling WDR77 with ab154190 at 1/50 dilution

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling WDR77 with ab154190 at 1/50 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded Human thyroid gland tissue using ab154190 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded Human normal testis tissue using ab154190 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded Human breast carcinoma tissue using ab154190 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of paraffin embedded Human thyroid gland carcinoma tissue using ab154190 showing +ve staining.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.