Epithelial Membrane Antigen (EMA) Antibody Staining Protocol for Immunohistochemistry


Description: Anti-EMA, E29 reacts with a variety of normal epithelial tissues including: secretory and ductal epithelial cells of the skin, breast and other organs of glandular differentiation including sebaceous, esophageal and gastric glands.


Primary Antibody

Name: EMA Antibody

Clone: E29, Mouse anti-Human

Supplier: Dako

Catalog Number: M0613

Dilution: 1:500 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 and plasma membrane
Images: Search image

Additional Information:
Tissue Type: Skin, Breast cancer, Colon cancer
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections, or cell smears
Positive Control: Skin, Breast cancer, Colon cancer
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


No need to perform antigen retrieval on frozen sections