The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Detects a band of approximately 47 kDa (predicted molecular weight: 43 kDa).
Use at an assay dependent concentration.
Regulates the activity of the neutrophil proteases elastase, cathepsin G, proteinase-3, chymase, chymotrypsin, and kallikrein-3.
Belongs to the serpin family. Ov-serpin subfamily.
Reactive bond 1 is specific for reaction with chymotrypsin-like protease such as cathepsin G, chymotrypsin or chymase, while reactive bond 2 is specific for reaction with elastase-like protease such as neutrophyl elastase, proteinase-3, pancreatic elastase or PSA.
ab47731 (2µg/ml) staining Serpin B1 in human spleen using an automated system (DAKO Autostainer Plus). Using this protocol there is cytoplasmic staining in both red and white pulp. Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 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 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, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.