CD68 Antibody Staining Protocol for Immunohistochemistry


Description: CD68 is a 110 kD transmembrane glycoprotein which is highly expressed in human monocytes including tissue macrophages and was identified to be a member of a group of hematopoietic molecules including CD43, CD34 and the lymph node ligand for L-selectin GlyCAM-1.1 The antigen is thought to play a role in endocytosis and/or lysosomal traffic.

Primary Antibody

Name: CD68, Macrophage Antibody

Clone: KP11, Mouse anti-Human

Supplier: Dako

Catalog Number: M0814

Dilution: 1:2000 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 min/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: Cytoplasmic
Images: Search image

Additional Information:
Tissue Type: Skin
Fixation: Formalin fixed paraffin sections
Positive Control: All tissue macrophages such as kidney, pancreas, skin, etc.
Negative Control: Omit primary antibody, isotype control, 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



1. Kunisch E., et al (2003) Macrophage specificity of three anti-CD68 monoclonal antibodies (KP1, EBM11, and PGM1) widely used for immunohistochemistry and flow cytometry. Ann Rheum Dis. 63(7):774-84.


2. Caffo M., et al (2005) CD68 and CR3/43 immunohistochemical expression in secretory meningiomas. Neurosurgery. 57(3):551-7; discussion 551-7. PubMed Abstract


3. Li WV, et al (1996) Immunohistochemical characterization of mast cell disease in paraffin sections using tryptase, CD68, myeloperoxidase, lysozyme, and CD20 antibodies. Mod Pathol. 9(10):982-8. PubMed Abstract