CD30 Antibody Staining Protocol for Immunohistochemistry


Description: This antibody labels anaplastic large-cell lymphoma (ALCL) and Reed-Sternberg cells, and is a useful tool for the identification of ALCL and as a secondary marker for Hodgkin’s disease. CD30 is a transmembrane cytokine receptor belonging to the tumor necrosis factor (TNF) receptor superfamily. Mature CD30 has a molecular mass of 120 kDa and is derived from a 90 kDa precursor protein. The extracellular domain of CD30 is homologous to that of TNF receptor superfamily members, whereas there is no homology in the cytoplasmic domain, suggesting major differences in signalling mechanisms. The intracellular part of CD30 possesses kinase activity, indicating that CD30 plays a role in regulating the function, differentiation and/or proliferation of normal lymphoid cells. A soluble 85 kDa form of CD30, sCD30, released from the membrane-bound molecule by proteolytic cleavage, can be detected in the sera of patients with CD30-expressing neoplasms. CD30 expression is found on Hodgkin and Reed-Sternberg (H-RS) cells, anaplastic large-cell lymphoma cells, and on activated B and T lymphocytes. In non-lymphoid tissues and neoplasms, CD30 expression has been confirmed in embryonal carcinomas, seminomas, decidual cells and mesotheliomas.


Primary Antibody

Name: CD30 Antibody

Clone: Ber H2, Mouse anti-Human

Supplier: Dako

Catalog Number: M0751

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: Membrane/cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Tonsil, thymus, spleen
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


Not need to perform antigen retrieval for acetone fixed frozen sections