General description
Filaggrin (UniProt: P20930; also known as FLG) is encoded by the FLG gene (Gene ID: 2312) in human. Filaggrin, a calcium-binding protein, is synthesized as a large, insoluble, highly phosphorylated precursor with several tandem copies of 324 amino acids in cytoplasmic granules. During terminal differentiation it undergoes dephosphorylation and proteolytic cleavage to remove linker peptides and produce functional basic filaggrin units. It is widely expressed and in skin tissue it is detected mainly in stratum granulosum of the epidermis. During terminal differentiation of mammalian epidermis, it aggregates keratin intermediate filaments and promotes disulfide-bond formation among the intermediate filaments. This creates the intracellular fibrous matrix of the cornified cells. Deimination of arginine to citrulline by peptidylarginine deiminase is reported to result in reduction in its affinity for cytokeratins. The N-terminal region of the mature filaggrin is shown to be heterogenous and is blocked by the formation of pyroglutamate. Clone AHF3 detects both the profilaggrin and filaggrin and exhibits reactivity in various immunochemical applications. (Ref.: Simon, M., et al. (1996). Biochem. J. 317(pt 1); 173 177; Simon, M., et al. (1995). Clin. Exp. Immunol. 100(1); 90-98; Simon, M., et al. (1995). J. Invest. Dermatol. 105(3); 432-437).
Immunogen
Partially purified Filaggrin from human breast skin tissue.
Application
Quality Control Testing
Evaluated by Immunocytochemistry in HaCat cells.
Immunocytochemistry Analysis: A 1:1,000 dilution of this antibody detected Filaggrin in HaCat cells.
Tested Applications
Immunohistochemistry Applications: A representative lot detected Filaggrin in Immunohistochemistry application.. (Henry, J., et al. (2011). FASEB J. 25(5):1567-76; Hsu, C-Y., et al. (2011). J Biol Chem. 286(26):23222-33).
Western Blotting Analysis: A representative lot detected Filaggrin in Western Blotting application. (Simon, M., et al. (1995). J Invest Dermatol. 105(3):432-7; Simon, M., et al. (1996). Biochem J. 317(Pt1)(Pt1):173-7; Henry, J., et al. (2011). FASEB J. 25(5):1567-76; Hsu, C-Y., et al. (2011). J Biol Chem. 286(26):23222-33; Pellerin, L., et al. (2013). J Allergy Clin Immunol. 131(4):1094-102; Hsu, C-Y., et al. (2017). J Invest Dermatol. 137(2):422-429).
Electron Microscopy: A representative lot detected Filaggrin in Electron Microscopy application. (Simon, M., et al. (1995). J Invest Dermatol. 105(3):432-7; Henry, J., et al. (2011). FASEB J. 25(5):1567-76).
Immunofluorescence Analysis: A representative lot detected Filaggrin in Immunofluorscence application. (Simon, M., et al. (1995). J Invest Dermatol. 105(3):432-7; Henry, J., et al. (2011). FASEB J. 25(5):1567-76; Hsu, C-Y., et al. (2011). J Biol Chem. 286(26):23222-33).
ELISA Analysis: A representative lot detected Filaggrin in ELISA application. (Simon, M., et al. (1995). Clin Exp Immunol. 100(1):90-8; Simon, M., et al. (1996). Biochem J. 317(Pt1)(Pt1):173-7; Miedouge, M., et al. (2001). Tumour Biol. 22(1):19-26).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone AHF3 is a mouse monoclonal antibody that detects Filaggrin and Profilaggrin.
Physical form
Purified mouse monoclonal antibodyIgG1κ in buffer containing PBS with 0.1% sodium azide with 50% glycerol.
Preparation Note
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
