All lanes : Anti-Kv4.2/KCND2 antibody [K57/1] (ab192762) at 2 µg/ml

Lane 1 : Mouse Brain normal tissue lysate
Lane 2 : Rat Brain normal tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : HRP-conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 70 kDa
Additional bands at: 55 kDa (possible IgG)


Exposure time: 20 minutes

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab192762 overnight at 4°C. Antibody binding was detected using a goat anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

IHC image of Kv4.2/KCND2  staining in a section of formalin-fixed paraffin-embedded normal mouse brain 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 20mins. The section was then incubated with ab192762, 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.

IHC image of Kv4.2/KCND2 staining in a section of formalin-fixed paraffin-embedded normal rat brain 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 20mins. The section was then incubated with ab192762, 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.