All lanes : Anti-Perilipin-1 antibody (ab126639) at 1 µg/ml

Lane 1 : P0 Mouse Brown Adipose Tissue Mouse Tissue Lysate
Lane 2 : P7 Rat Brown Adipose Tissue Rat Tissue Lysate

Lysates/proteins at 10 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: 56 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?


Exposure time: 2 minutes

The predicted molecular weight of Perilipin-1 is 56 kDa (SwissProt), however we expect to observe a banding pattern around 62 kDa.

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 5% Bovine Serum Albumin before being incubated with ab126639 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

IHC image of Perilipin-1 staining in human breast adipose formalin fixed paraffin embedded tissue, 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 ab126639, 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 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.