The antibody recognizes endogenous NCF1/p47-phox protein exclusively when phosphorylated at Ser304, a key site for activating the NADPH oxidase complex . Phosphorylation of Ser304 by PRKCD (protein kinase C delta) induces structural changes in NCF1, enabling its translocation to the cell membrane and subsequent assembly of the functional oxidase complex . This specificity ensures precise detection of activated NCF1 in experimental systems, avoiding cross-reactivity with non-phosphorylated forms .
The antibody supports a range of assays, including:
| Application | Dilution Range | Source |
|---|---|---|
| Western Blot (WB) | 1:500–1:2000 | |
| Immunohistochemistry (IHC) | 1:100–1:300 | |
| Immunofluorescence (IF) | 1:50–200 | |
| ELISA | 1:20000 | |
| Immunocytochemistry (ICC) | Not specified |
Validation data include immunohistochemical staining of human lung cancer tissue and Western blot analysis of COS7 and A549 cells treated with UV . The antibody demonstrates high sensitivity and specificity, with blocking peptide controls confirming target engagement .
NCF1/p47-phox is a critical organizer of the NADPH oxidase complex, which generates superoxide radicals in phagocytes . Dysregulation of NCF1 phosphorylation is implicated in chronic granulomatous disease (CGD), an immunodeficiency characterized by recurrent infections due to impaired oxidative burst . This antibody enables researchers to: