Epithelial Membrane Antigen (EMA) Antibody Staining Protocol for Immunohistochemistry

 

Description: Epithelial membrane antigen (EMA), is a mucin-like glycoprotein. Antibody to EMA has been shown useful as a pan- epithelial marker for detecting early metastatic loci of carcinoma in the bone marrow or liver.

 

Primary Antibody

Name: EMA Ab-3 Antibody

Clone: E29, Mouse anti-Human

Supplier: Lab Vision

Catalog Number: MS-741

Dilution: 1:50 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: No antigen retrieval is needed.

Buffer/pH value: N/A

Heat/Cool Temperature: N/A

Heat/Cool Time: N/A

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 and cell membrane

Images: Search image

Additional Information:

Tissue Type: Breast cancer, Colon carcinoma

Fixation: Formalin fixed paraffin sections

Positive Control: Breast cancer, Colon 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

Notes:

Not tested on frozen sections

 

References: