Anti-PC4 antibody (ab72132)

All lanes : Anti-PC4 antibody (ab72132) at 0.04 µg/ml

Lane 1 : Whole cell lysate from Hela cells at 50 µg
Lane 2 : Whole cell lysate from Hela cells at 15 µg
Lane 3 : Whole cell lysate from Hela cells at 5 µg

Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Additional bands at: 190 kDa. We are unsure as to the identity of these extra bands.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling PC4 with ab72132 at 1/200 (1µg/ml). Detection: DAB.

Immunoprecipitation/ Western Blot of PC4.
Lane 1: ab72132 at 3µg/mg whole cell lysate.
Lane 2: Control IgG.

ab72132 at 0.1µg/ml for WB.
Whole cell lysate from Hela cells at 1mg for IP, 20% of IP loaded.
Detection:Chemiluminescence with an exposure time of 30 seconds.

IHC image of ab72132 staining in human normal pancreas formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM 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 ab72132, 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.