CD16 Antibody Staining Protocol for Immunohistochemistry


Description: CD16 is a low affinity Fc receptor of complexed IgG, Fc/gamma RIII and expressed on NK cells, granulocytoes, activated macrophages and a subset of T cells. CD16 exists both as a glycosyl-phosphatidylinositol-anchored protein in polymorphonuclear cells and as a transmembrane protein in NK cells. This antigen will help detect CD16 and NK cells in a variety of tissues.


Primary Antibody

Name: CD16 Antibody

Clone: 2H7, Mouse anti-Human

Supplier: Vector Labs

Catalog Number: VP-C333

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: 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 staining of macrophage, neutrophils, NK cells and granulocytes
Images: Search image

Additional Information:
Tissue Type: Tonsil
Fixation: Formalin fixed paraffin sections
Positive Control: Tonsil
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 effective on fixed frozen sections