All lanes : Anti-AACT antibody (ab9374) at 1 µg/ml

Lane 1 : Human spinal cord tissue lysate - total protein (ab29188)
Lane 2 : Human colon tissue lysate - total protein (ab30051)

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 48 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Standard Curve for AACT (Analyte: AACT protein (Human) (ab80517) ); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal to AACT (ab54693) at 5µg/ml and Detector Antibody Rabbit polyclonal to AACT (ab9374) at 0.5µg/ml.

Human normal skin. Staining is mainly membrane-bound and cytoplasmic. Left panel: with primary antibody at 2 ug/ml. Right panel: negative control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature: sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffers citrate EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for rabbit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.