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pMD18-T Simple Vector is a high-efficiency TA cloning vector constructed from pUC18, of which the initial multiple cloning sites (MCS) were destroyed. Thus the cDNA should be amplified by PCR with primers containing a restriction site for subclone. Competent cells appropriate for pUC18 are also appropriated for the Vector, e.g. JM109, DH5α, TOP10. The pMD18-T Simple Vector is 2.6kb in size. Selection of the plasmid in E. coli is conferred by the ampicillin resistance gene. The coding sequence was inserted by TA cloning at site 425.
The coding sequence can be amplified by PCR with M13-47 and RV-M primers.
|Human ApoE ORF mammalian expression plasmid, C-GFPSpark tag||HG10817-ACG|
|Human ApoE ORF mammalian expression plasmid, C-OFPSpark / RFP tag||HG10817-ACR|
|Human ApoE ORF mammalian expression plasmid, C-Flag tag||HG10817-CF|
|Human ApoE ORF mammalian expression plasmid, C-His tag||HG10817-CH|
|Human ApoE ORF mammalian expression plasmid, C-Myc tag||HG10817-CM|
|Human ApoE ORF mammalian expression plasmid, C-HA tag||HG10817-CY|
|Human ApoE ORF mammalian expression plasmid, His tag||HG10817-M-H|
|Human ApoE ORF mammalian expression plasmid, N-Flag tag||HG10817-NF|
|Human ApoE ORF mammalian expression plasmid, N-His tag||HG10817-NH|
|Human ApoE ORF mammalian expression plasmid, N-Myc tag||HG10817-NM|
|Human ApoE ORF mammalian expression plasmid, N-HA tag||HG10817-NY|
|Human ApoE natural ORF mammalian expression plasmid||HG10817-UT|
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Apolipoprotein E (ApoE) is a 34.2 kDa glycosylated protein with 299 amino acid residues. There are three isoforms in human (apoE2, apoE3, and apoE4) due to different amino acid residues at positions 112 and 158. ApoE is synthesized predominantly in the liver, but also by cells in the spleen, brain, lung, kidney, ovary, adrenal, and muscle tissues. Hepatic parenchyma cells are the main apoE producing cells in mammalian body, probably accounting for two thirds to three fourths of the plasma apoE . In the nervous system, apoE mRNA is present in neurons, astrocytes, ependymal cells, nonmyelinating Schwann cells, but not in microglia, oligodendroglia, choroidal cells, or myelinating Schwann cells. ApoE produced by mammalian cells exists in different forms, monomers, dimers, modified, unmodified, lipid-rich, and lipid-poor, and so forth. ApoE plays a double-role in immune responses. Both apoE containing lipoproteins and multimers of synthetic apoE peptides inhibited proliferation of cultured lymphocytes by inhibiting DNA synthesis and reducing phospholipid turnover in T cells. ApoE can also affect innate and acquired immune responses in vitro by its ability to suppress stimulation of cultured neutrophils. ApoE can bind lipopolysaccharide (LPS), attenuate the inflammatory response, and thus reduce LPS induced lethality. Injection of LPS stimulated higher expression of inflammatory cytokines like interleukin (IL)-1β, IL-12, and interferon-γ (IFN-γ), as well as IL-6.