|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.
|Human KRAS ORF mammalian expression plasmid, C-GFPSpark tag||HG12259-ACG|
|Human KRAS ORF mammalian expression plasmid, C-OFPSpark / RFP tag||HG12259-ACR|
|Human KRAS ORF mammalian expression plasmid, N-GFPSpark tag||HG12259-ANG|
|Human KRAS ORF mammalian expression plasmid, C-Flag tag||HG12259-CF|
|Human KRAS ORF mammalian expression plasmid, C-His tag||HG12259-CH|
|Human KRAS ORF mammalian expression plasmid, C-Myc tag||HG12259-CM|
|Human KRAS ORF mammalian expression plasmid, C-HA tag||HG12259-CY|
|Human KRAS Gene cDNA clone plasmid||HG12259-G|
|Human KRAS ORF mammalian expression plasmid, N-Flag tag||HG12259-NF|
|Human KRAS ORF mammalian expression plasmid, N-His tag||HG12259-NH|
|Human KRAS ORF mammalian expression plasmid, N-Myc tag||HG12259-NM|
|Human KRAS ORF mammalian expression plasmid, N-HA tag||HG12259-NY|
|Human KRAS natural ORF mammalian expression plasmid||HG12259-UT|
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K-Ras belongs to the small GTPase superfamily, Ras family. As other members of the Ras family, K-Ras is a GTPase and is an early player in many signal transduction pathways. It is usually tethered to cell membranes because of the presence of an isoprenyl group on its C-terminus. K-Ras functions as a molecular on/off switch. Once it is turned on it recruits and activates proteins necessary for the propagation of growth factor and other receptors' signal, such as c-Raf and PI 3-kinase. It binds to GTP in the active state and possesses an intrinsic enzymatic activity which cleaves the terminal phosphate of the nucleotide converting it to GDP. Upon conversion of GTP to GDP, K-Ras is turned off. The rate of conversion is usually slow but can be sped up dramatically by an accessory protein of the GTPase activating protein class, for example RasGAP. In turn K-Ras can bind to proteins of the Guanine Nucleotide Exchange Factor class, for example SOS1, which forces the release of bound nucleotide. Subsequently, K-Ras binds GTP present in the cytosol and the GEF is released from ras-GTP. Besides essential function in normal tissue signaling, the mutation of a K-Ras gene is an essential step in the development of many cancers. Several germline K-Ras mutations have been found to be associated with Noonan syndrome and cardio-facio-cutaneous syndrome. Somatic K-Ras mutations are found at high rates in Leukemias, colon cancer, pancreatic cancer and lung cancer.