All lanes : Anti-FMRP (phospho S499) antibody (ab48127) at 2 µg/ml

Lane 1 : Human brain tissue lysate - total protein (ab29466)
Lane 2 : Mouse Brain Tissue Lysate

Lysates/proteins at 20 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: 71 kDa
Additional bands at: 71 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 8 minutes

Serially diluted ab48127 was bound to immobilised phospho (ab48658) - or control (unmodified) peptides (1 microgram x mL^-1). The antibody was detected by HRP-labelled goat anti-rabbit IgG (ab97080; diluted 50,000 times) and signal was developed with TMB substrate

IHC image of FMRP (phospho S499) staining in human cerebral cortex 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 48127, 1µg/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.

All lanes : Anti-FMRP (phospho S499) antibody (ab48127) at 0.25 µg/ml

Lane 1 : Marker
Lane 2 : Zebrafish brain homogenate at 20 µg
Lane 3 : Zebrafish heart homogenate at 20 µg

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 71 kDa
Observed band size: 71 kDa


Exposure time: 5 minutes