|Datasheet||Specific References||Reviews||Related Products||Protocols|
|Vector Type||Mammalian Expression Vector|
|Expression Method||Constiutive, Stable / Transient|
|Selection In Mammalian Cells||Hygromycin|
FLAG-tag, or FLAG octapeptide, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology. It can be used for affinity chromatography, then used to separate recombinant, overexpressed protein from wild-type protein expressed by the host organism. It can also be used in the isolation of protein complexes with multiple subunits.
A FLAG-tag can be used in many different assays that require recognition by an antibody. If there is no antibody against the studied protein, adding a FLAG-tag to this protein allows one to follow the protein with an antibody against the FLAG sequence. Examples are cellular localization studies by immunofluorescence or detection by SDS PAGE protein electrophoresis.
The peptide sequence of the FLAG-tag from the N-terminus to the C-terminus is: DYKDDDDK (1012 Da). It can be used in conjunction with other affinity tags, for example a polyhistidine tag (His-tag), HA-tag or Myc-tag. It can be fused to the C-terminus or the N-terminus of a protein. Some commercially available antibodies (e.g., M1/4E11) recognize the epitope only when it is present at the N-terminus. However, other available antibodies (e.g., M2) are position-insensitive.
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-GFPSpark tag||MG50789-ACG|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-OFPSpark / RFP tag||MG50789-ACR|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-Flag tag||MG50789-CF|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-His tag||MG50789-CH|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-Myc tag||MG50789-CM|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, C-HA tag||MG50789-CY|
|Mouse RAC1 / MIG5 Gene cDNA clone plasmid||MG50789-G|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, N-Flag tag||MG50789-NF|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, N-His tag||MG50789-NH|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, N-Myc tag||MG50789-NM|
|Mouse RAC1 / MIG5 ORF mammalian expression plasmid, N-HA tag||MG50789-NY|
|Mouse RAC1 / MIG5 natural ORF mammalian expression plasmid||MG50789-UT|
|Learn more about expression Vectors|
RAC1 is a GTPase which belongs to the RAS superfamily of small GTP-binding proteins. Members of this superfamily appear to regulate a diverse array of cellular events, including the control of cell growth, cytoskeletal reorganization, and the activation of protein kinases. Two transcript variants encoding different isoforms have been found for RAC1 gene. RAC1 is a plasma membrane-associated small GTPase which cycles between active GTP-bound and inactive GDP-bound states. In its active state, binds to a variety of effector proteins to regulate cellular responses such as secretory processes, phagocytosis of apoptotic cells, epithelial cell polarization and growth-factor induced formation of membrane ruffles. RAC1 p21/rho GDI heterodimer is the active component of the cytosolic factor sigma 1, which is involved in stimulation of the NADPH oxidase activity in macrophage. RAC1 is essential for the SPATA13-mediated regulation of cell migration and adhesion assembly and disassembly. RAC1's isoform B has an accelerated GEF-independent GDP/GTP exchange and an impaired GTP hydrolysis, which is restored partially by GTPase-activating proteins. It is able to bind to the GTPase-binding domain of PAK but not full-length PAK in a GTP-dependent manner, suggesting that the insertion does not completely abolish effector interaction.
Stat3 is an important transcription factor that regulates both proinflammatory and anit-apoptotic pathways in the heart. It forms a multiprotein complex with RAC1 and PKC in an H/R-dependent manner by expression of constitutively active Rac1 mutant protein, and by RNA silencing of RAC1. Selective inhibition of PKC with calphostin C produces a marked suppression of Stat3 S727 phosphorylation. The association of Stat3 with Rax1 occurs predominantly at the cell membrane, but also inside the nucleus, and occurs through the binding of the coiled-coil domain of Stat3 to the 54 NH(2)-terminal residues of RAC1. Transfection with a peptide comprising the NH(2)-terminal 17 amino acid residues of RAC1-dependent signaling pathways resulting in physical association between Rac1 and Stat3 and the formation of a novel multiprotein complex with PKC.