Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: ROR2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: MCF7 whole cell lysate (20 µg)
Lane 4: U20S whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab190145 observed at 125 kDa. Red - loading control, ab181602, observed at 37 kDa.

ab190145 was shown to recognize ROR2 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when ROR2 knockout samples were examined. Wild-type and ROR2 knockout samples were subjected to SDS-PAGE. Ab190145 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) ab216772 and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) ab216777 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes : Anti-ROR2 antibody [Nt 2535-2835] (ab190145) at 1 µg/ml

Lane 1 : Kidney (Mouse) Tissue Lysate
Lane 2 : Lung (Rat) Tissue Lysate
Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 105 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?


Exposure time: 2 minutes

IHC image of ROR2 staining in Normal Human Stomach formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F/B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab190145, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

IHC image of ROR2 staining in Human normal prostate formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab190145, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre