CD163 Antibody Staining Protocol for Immunohistochemistry


Description: CD163 is cell surface glycoprotein, a 175kD molecule also known as ED2. CD163 is expressed by approximately 50% of peritoneal macrophages, a subset of splenic macrophages, and by macrophages in most other tissues. However, it is not expressed by monocytes, alveolar macrophages or microglial cells.


Primary Antibody

Name: CD163 (EDHu-1) Antibody

Clone: EDHu-1, Mouse anti-Human

Supplier: AbD Serotec

Catalog Number: MCA1853

Dilution: 1:400 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: No pretreatment needed. IHC-TekTM Epitope Retrieval Solution (Cat# IW-1100) may improve staining intensity.
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/Cytoplasm
Images: Search image

Additional Information:
Species Reactivity: Human
Tissue Type: Human spleen
Fixation: Formalin fixed paraffin sections, or acetone fixed frozen sections
Positive Control: Human 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 on acetone fixed frozen sections