All lanes : Anti-Adrenomedullin/ADM antibody (ab69117) at 1 µg/ml

Lane 1 : Human breast tissue lysate - total protein (ab30090)
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 3 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
Lane 4 : MDA-MB-361 (Human breast adenocarcinoma cell line) Whole Cell Lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 20 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?
Additional bands at: 170 kDa, 70 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 8 minutes

IHC image of Adrenomedullin/ADM staining in Human placenta formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab69117, 5µg/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.