Detects Furin from canine and mouse cells as well as transfected human Furin.
This antibody does not detect endogenous Furin from BSC-40, HeLa, J774A.1 BPAEC, or CHO cells nor from rat skeletal muscle, spleen, kidney, ovary, testes, heart, or brain tissue.
Furin is a membrane-associated, calcium-dependent, serine protease that belongs to the subtilisin-like prohormone convertase (PC) family. Members of this family of cellular enzymes cleave most prohormones and neuropeptide precursors. Numerous other cellular proteins, some viral proteins, and bacterial toxins that are transported by the constitutive secretory pathway are also targeted for maturation by PCs. Furin and other PC family members share structural similarities which include a heterogeneous ~10 kDa amino-terminal proregion, a highly conserved ~55 kDa subtilisin-like catalytic domain, and carboxyl-terminal domain that is heterogeneous in length and sequence. These enzymes become catalytically active following proregion cleavage within the appropriate cellular compartment.
Furin is the only known PC to possess a transmembrane domain. Cleavage of target proteins occurs at the carboxyl-terminus of the furin consensus sequence, RX(K/R)R. It has been shown that the acidic peptide sequence, C771PSDSEEDEG780, localizes furin to the trans-Golgi-network. Phosphorylation of serine residues within this region modulates intracellular routing of Furin protein. An additional signaling domain includes the tetrapeptide sequence, Y759KGL762, which directs internalization from the cell surface.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use at an assay dependent concentration.
Use at an assay dependent concentration. PubMed: 18174282
Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 87 kDa).Can be blocked with Furin peptide (ab4989).
Representing Furin in BSC-40 cells transfected with the human Furin gene.
Use at an assay dependent concentration. PubMed: 23418414
Furin is likely to represent the ubiquitous endoprotease activity within constitutive secretory pathways and capable of cleavage at the RX(K/R)R consensus motif.
Seems to be expressed ubiquitously.
Belongs to the peptidase S8 family. Furin subfamily. Contains 1 homo B/P domain.
Contains a cytoplasmic domain responsible for its TGN localization and recycling from the cell surface.
The inhibition peptide, which plays the role of an intramolecular chaperone, is autocatalytically removed in the endoplasmic reticulum (ER) and remains non-covalently bound to furin as a potent autoinhibitor. Following transport to the trans Golgi, a second cleavage within the inhibition propeptide results in propeptide dissociation and furin activation. Phosphorylation is required for TGN localization of the endoprotease. In vivo, exists as di-, mono- and non-phosphorylated forms.
Golgi apparatus > trans-Golgi network membrane. Cell membrane. Shuttles between the trans-Golgi network and the cell surface. Propeptide cleavage is a prerequisite for exit of furin molecules out of the endoplasmic reticulum (ER). A second cleavage within the propeptide occurs in the trans Golgi network (TGN), followed by the release of the propeptide and the activation of furin.
The band observed at 63 kDa could potentially be a cleaved form of Furin due to the presence of a 24 amino acid signal peptide and a 83 amino acid propeptide.
Immunocytochemistry/ Immunofluorescence - Anti-Furin antibody (ab3467)Image courtesy of an anonymous Abreview.
ab3467 staining Furin in human A549 cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized using 0.1% Triton X-100, blocked with 1% BSA/ 5%FBS/PBS for 1 hour at 25°C and then incubated with ab3467 at a 1/100 dilution for 1 hour at 25°C. The secondary used was an Alexa-Fluor 546 conjugated goat anti-rabbit polyclonal used at a 1/500 dilution. Cells were counterstained with DAPI (436nM, 5 mins) and Phalloidin-633 (1unit/coverslip) and mounted with prolong gold. No staining seen in secondary control and furin staining was punctate and throughout the cytoplasm.