The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/20 - 1/40. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol. Note: The use of H2O2 to block endogenous peroxidase has a detrimental effect on the epitope recognised by ab67001. Solutions containing 1% or greater H2O2 can dramatically REDUCE the level of staining achievable. Blocking after antigen retrieval can also lead to negative results. Therefore, it is recommended that endogenous peroxidase should be blocked with 0.5% H2O2/methanol for 10 minutes BEFORE antigen retrieval. If this recommendation is not followed, staining intensity may be reduced.
FunctionAccessory protein for MHC class-II antigen/T-cell receptor interaction. May regulate T-cell activation. Induces the aggregation of lipid rafts.
Post-translational modificationsPalmitoylation and association with LCK contribute to the enrichment of CD4 in lipid rafts.
Cellular localizationCell membrane. Localizes to lipid rafts. Removed from plasma membrane by HIV-1 Nef protein that increases clathrin-dependent endocytosis of this antigen to target it to lysosomal degradation. Cell surface expression is also down-modulated by HIV-1 Envelope polyprotein gp160 that interacts with, and sequesters CD4 in the endoplasmic reticulum.