The monoclonal antibody 6D8 recognizes the extracellular domain of human desmoglein-2. Desmogleins and desmocollins are members of the cadherin family of transmembrane proteins that together make up the core of the desmosome, a structure that provides transmembrane strength to tissues undergoing mechanical stress. The desmosomal cadherins, desmogleins and desmocollins, mediate calcium-dependent cell-cell adhesion by forming homotypic and heterotypic interactions with one another. The multiprotein desmosomal complex also includes the cytoplasmic desmosomal plaque proteins plakoglobin, phakophilins, and desmoplakin, which bind to the intracellular domain of the desmogleins and function to anchor the keratin intermediate filament network to site of cellcell contacts.
In human, four desmogleins have been identified (Dsg14). Desmogleins are synthesized with a signal peptide that directs them to the endoplasmic reticulum and a proregion that is removed during protein processing. The mature protein includes four highly conserved extracellular domains (EC 14) and a fifth membrane proximal, more variable EC domain that is referred to as the “extracellular anchor domain. Desmoglein-2 is expressed on various cells including simple epithelia and myocardium, tumors and and many cell cultures.
Desmogleins play critical roles in cell adhesion and skin blistering diseases, as they are the target antigens of autoimmune antibodies and bacterial toxins. Desmosomal dysfunction has been implicated in a number of diseases, including striate palmoplantar keratoderma, skin fragility, and ectodermal dysplasia, and most recently arrhythmic right ventricular cardiomyopathy (ARVC).
Frozen sections, Immuno fluorescence, Immuno precipitation, Western blot
W: A reduced sample treatment (Laemmli buffer) and SDS-PAGE was used. The band size is ~165 kDa (Ref.5).
IHC-F:- Tissues frozen in OCT were fixed in 100% methanol for 15 min at 20 °C, permeabilized in 50:50 methanol/acetone for 2 min at 20 °C, and then treated with 1% Triton X-100 in PBS for 5 min at room temperature. Non-specific sites were blocked for 30min in blocking buffer (PBS/0.1% Triton X-100/1% BSA); (Ref.5).
For immune fluorescence and Western blotting, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:10.