Description:
Monoclonal Anti-Epithelial Antigen, Clone Ber-EP4 (Anti-epithelial
antigen) reacts with the peptide portion of a 34 kDa and 39 kDa
glycoprotein expressed on the membrane and in the cytoplasm of
normal epithelial cells and carcinomas of various origins. In
immunoprecipitation studies, antiepithelial antigen has been shown
to precipitate a 34 kDa and a 39 kDa polypeptide from radiolabelled
MCF-7 cells. Under non-reducing conditions the molecular weight was
found to be slightly higher (39 kDa and 44 kDa). Deglycosylation of
the precipitate reduced the molecular weight to 31 kDa and 36 kDa
respectively. Internal labelling of MCF-7 and K562 cells for up to
24 hours resulted in the precipitation of only one chain (39 kDa)
after two hours of labelling, suggesting a 39 kDa gene product for
both chains. Anti-epithelial antigen, Clone Ber-EP4 has been shown
to be equivalent in specificity to monoclonal antibody HEA15.1-3
Sequential immunoprecipitation and cross-blocking experiments
demonstrated that both antibodies recognize the same antigen.
Primary Antibody
Name: Epithelial Antigen Antibody |
Clone: Ber EP4, Mouse anti-Human |
Supplier: Dako |
Catalog Number: M0804 |
Dilution: 1:20 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 |
Device: IHC-TekTM Epitope Retrieval Steamer Set (Cat# IW-1102) |
Buffer/pH value: IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) or IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
Heat/Cool Temperature: 95 ºC - 100 ºC/room temperature |
Heat/Cool Time: 20 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Membrane/cytoplasmic |
Images: Search image |
Tissue Type: Pancreas, pituitary gland, small intestine, colon, kidney. |
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections |
Positive Control: Pancreas, pituitary gland, small intestine, colon, kidney. |
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 |
References: