ARFIP1 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

ADP-ribosylation factors, or ARFs, enhance the ADP ribosyltransferase activity of cholera toxin and are implicated in vesicle transport between endoplasmic reticulum and the Golgi complex. Arfaptin 1 is recruited from the cytosol to Golgi membranes by ARFs in a guanosine 5'-O-(3-thiotriphosphate)-dependent and brefeldin A-sensitive manner but is not a constituent of coatomer. Arfaptin 1 binds to nonmyristoylated GTP-bound ARF3, but not to GDP-bound ARF3, and also to ARF1, another class I ARF. It binds with lower affinity to ARF5, a class II ARF, and with very little affinity to ARF6, a class III ARF. POR1 (also designated arfaptin 2) was first isolated as a Rac 1 binding protein necessary for Rac mediated Actin polymerization and the subsequent formation of membrane ruffles and lamellipodia. POR1 has also been shown to interact with the ADP ribosylation factor ARF6, a GTPase that associates with the plasma membrane and intracellular endosome vesicles, in a GTP dependent manner. The association of POR1 with ARF6 stimulates induction of Actin polymerization. POR1 appears to play a regulatory role through multiple signaling pathways in the reorganization of the cytoskeletal structure.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-07334

• ARFIP1 Antibodies
• ARFIP1 Lysates
• ARFIP1 Proteins