1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1/100 - 1/250.
Human IgA (immunoglobulin A) is a glycosylated protein of 160 kDa and is produced as a monomer or as a J chain linked dimer. Monomeric IgA constitutes 5-15 % of the serum immunoglobulins whereas dimeric IgA is localized to mucosa surfaces such as saliva, gastrointestinal secretion, bronchial fluids and milk. Mucosal IgA plays a major role in host defence by neutralising infectious agents at mucosal surfaces. The production is usually local and antigen specific IgA producing B cells can be found in regions under the lamina propria where they mature into dimeric IgA producing plasma cells. IgA deficiency is the most common immunodeficiency that may affect both serum and mucosal produced IgA.
The secretory component is a component of immunoglobulin A (IgA) which consists of a portion of the polymeric immunoglobulin receptor. Polymeric IgA binds to the polymeric immunoglobulin receptor on the basolateral surface of epithelial cells and is taken up into the cell via transcytosis. The receptor-IgA complex passes through the cellular compartments before being secreted on the luminal surface of the epithelial cells, still attached to the receptor. Proteolysis of the receptor occurs and the dimeric IgA molecule, along with the secretory component, are free to diffuse throughout the lumen.
Capture ab 1:60 dilution (1μg in 1mg lysate). Primary ab for WB ab124716 at 1:1000 dilution (0.18μg/ml). Secondary ab Anti-Rabbit IgG (HRP), specific to the non5% NFDM/TBST -reduced form of IgG. Secondary ab concentration 1:1500 dilution. Blocking buffer and concentration 5% NFDM/TBST. Diluting buffer and concentration 5% NFDM/TBST Lane 1 (+) Human plasma Lane 2 (-) Rabbit monoclonal IgG (ab172730) instead of ab124716 in Human plasma
All lanes : Anti-IgA antibody [EPR5367-76] (ab124716) at 0.18 µg/ml (ab124716 at 1:1000 dilution (0.18µg/ml))
Lane 1 : Human plasma with NFDM/TBST Lane 2 : Rabbit monoclonal IgG (ab172730) instead of ab124716 in Human plasma with NFDM/TBST
Blocking peptides at 5 % per lane.
Secondary All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution