CA19-9 (Sialyl Lewis a) Antibody Staining Protocol for Immunohistochemistry


Description: The Lewis blood group antigens Lewisa and Lewisb are carbohydrates that form epitopes on glycolipids and glycoproteins. CA19-9 is an epitope on the sialylated Lewisa carbohydrate structure. Sialylated Lewisa plays a role in cell adhesion by acting as a functional ligand for the inducible adhesion molecule E-selectin.


Primary Antibody

Name: CA19-9 Antibody

Clone: C241:5:1:4, Mouse anti-Human

Supplier: Novocastra Labs

Catalog Number: NCL-CA19-9

Dilution: 1:200 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: 60min/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: 95-100 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: Extracellular membrane-associated
Images: Search image

Additional Information:
Tissue Type: Small bowel
Fixation: Formalin-fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Small bowel
Negative Control: Omit primary antibody, isotype control, 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


There is no need to perform antigen retrieval for acetone fixed frozen sections