HSA Antibody Staining Protocol for Immunohistochemistry


Description: Hepatocyte Specific Antigen (HSA) is a monoclonal antibody is considered almost specific for normal and neoplastic hepatocytes. It has been demonstrated consistently in the vast majority of hepatocellular carcinomas. Studies have shown the utility of HSA in the differential diagnosis of hepatacellular carcinimoa, cholangiocarcinoma and hepatoblastomas. HSA can be used in a panel with CEA, CK7, AFP and CD10. Some focal staining of gastric biopsies and subpopulation of gastric carcinomas has been observed.


Primary Antibody

Name: HSA Antibody

Clone: OCH1E5, Mouse monoclonal

Supplier: Biocare Medical

Catalog Number: CM166A

Dilution: 1:100 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed.

Incubation Time/Temp: 60 minutes/room temperature

Antigen Retrieval
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102)
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100)
Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature
Heat/Cool Time: 20 minutes/20 minutes

Detection Methods
Standard Method: ABC Method or LSAB Method
Enhanced Method: Polymeric Methods

Chromogen Substrate
Reagent: DAB
Incubation Time/Temperature: 1-3 minutes/room temperature

Reagent: Mayer's Hematoxylin
Staining Time: 30 seconds

Staining Pattern: Cytoplasmic
Images: Search image

Additional Information:

Tissue Type: Liver

Fixation: Formalin fixed paraffin sections

Positive Control: Liver, liver carcinoma

Negative Control: Omit primary antibody, isotype control or absorption control

Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary


Not tested on frozen sections.