( We provide with CD4 qPCR primers for gene expression analysis, MP200164 )
|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 CD4 Gene ORF cDNA clone expression plasmid, C-GFPSpark tag||MG50134-ACG|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, C-OFPSpark tag||MG50134-ACR|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, C-Flag tag||MG50134-CF|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, C-His tag||MG50134-CH|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, C-Myc tag||MG50134-CM|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, C-HA tag||MG50134-CY|
|Mouse CD4 Gene ORF cDNA clone in cloning vector||MG50134-M|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, N-Flag tag||MG50134-NF|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, N-His tag||MG50134-NH|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, N-Myc tag||MG50134-NM|
|Mouse CD4 Gene ORF cDNA clone expression plasmid, N-HA tag||MG50134-NY|
|Mouse CD4 Gene ORF cDNA clone expression plasmid||MG50134-UT|
|Learn more about expression Vectors|
T-cell surface glycoprotein CD4, is a single-pass type I membrane protein. CD4 contains three Ig-like C2-type (immunoglobulin-like) domains and one Ig-like V-type (immunoglobulin-like) domain. CD4 is a glycoprotein expressed on the surface of T helper cells, regulatory T cells, monocytes, macrophages, and dendritic cells. The CD4 surface determinant, previously associated as a phenotypic marker for helper/inducer subsets of T lymphocytes, has now been critically identified as the binding/entry protein for human immunodeficiency viruses (HIV). The human CD4 molecule is readily detectable on monocytes, T lymphocytes, and brain tissues. All human tissue sources of CD4 bind radiolabeled gp120 to the same relative degree; however, the murine homologous protein, L3T4, does not bind the HIV envelope protein. CD4 is a co-receptor that assists the T cell receptor (TCR) to activate its T cell following an interaction with an antigen presenting cell. Using its portion that resides inside the T cell, CD4 amplifies the signal generated by the TCR. CD4 interacts directly with MHC class II molecules on the surface of the antigen presenting cell via its extracellular domain. The CD4 molecule is currently the object of intense interest and investigation both because of its role in normal T-cell function, and because of its role in HIV infection. CD4 is a primary receptor used by HIV-1 to gain entry into host T cells. HIV infection leads to a progressive reduction of the number of T cells possessing CD4 receptors.
Viral protein U (VpU) of HIV-1 plays an important role in downregulation of the main HIV-1 receptor CD4 from the surface of infected cells. Physical binding of VpU to newly synthesized CD4 in the endoplasmic reticulum is an early step in a pathway leading to proteasomal degradation of CD4. Amino acids in both helices found in the cytoplasmic region of VpU in membrane-mimicking detergent micelles experience chemical shift perturbations upon binding to CD4, whereas amino acids between the two helices and at the C-terminus of VpU show no or only small changes, respectively. Paramagnetic spin labels were attached at three sequence positions of a CD4 peptide comprising the transmembrane and cytosolic domains of the receptor. VpU binds to a membrane-proximal region in the cytoplasmic domain of CD4.