Standard Curve for alpha 1 Antichymotrypsin (Analyte: alpha 1 Antichymotrypsin protein (Human) (ab80517) ); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal to alpha 1 Antichymotrypsin (ab54693) at 5µg/ml and Detector Antibody Rabbit polyclonal to alpha 1 Antichymotrypsin (ab9374) at 0.5µg/ml.
All lanes : Anti-alpha 1 Antichymotrypsin antibody (ab9374) at 1 µg/mlLane 1 : Spinal Cord (Human) Tissue Lysate - adult normal tissue (ab29188)Lane 2 : Colon tissue lysate (Human) Tissue Lysate - adult normal tissue (ab30051)Lysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
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.
Anti-alpha 1 Antichymotrypsin antibody (ab9374) at 1 µg/ml + Human alpha 1 Antichymotrypsin full length protein (ab80517) at 0.01 µgSecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.Exposure time : 30 seconds