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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
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Name: HSA Antibody |
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Clone: OCH1E5, Mouse monoclonal |
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Supplier: Biocare Medical |
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Catalog Number: CM166A |
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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. |
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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 |
Counterstain
| Reagent: Mayer's Hematoxylin |
| Staining Time: 30 seconds |
Results:
| Staining Pattern: Cytoplasmic |
| Images: Search image |
Additional Information:
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Tissue Type: Liver |
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Fixation: Formalin fixed paraffin sections |
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Positive Control: Liver, liver carcinoma |
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Negative Control: Omit primary antibody, isotype control or absorption control |
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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 |
Notes:
Not tested on frozen sections. |
References: