|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.
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-GFPSpark 標籤||HG14439-ACG|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-OFPSpark 標籤||HG14439-ACR|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-GFPSpark 標籤||HG14439-ANG|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-OFPSpark 標籤||HG14439-ANR|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-Flag 標籤||HG14439-CF|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-His 標籤||HG14439-CH|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-Myc 標籤||HG14439-CM|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), C-HA 標籤||HG14439-CY|
|人 ABHD10 基因全長cDNA ORF(克隆載體)||HG14439-G|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-Flag 標籤||HG14439-NF|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-His 標籤||HG14439-NH|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-Myc 標籤||HG14439-NM|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體), N-HA 標籤||HG14439-NY|
|人 ABHD10 基因全長cDNA ORF克隆 (表達載體)||HG14439-UT|
Mycophenolic acid (MPA), the active metabolite of the immunosuppressant mycophenolate mofetil (MMF), is primarily metabolized by glucuronidation to a phenolic glucuronide (MPAG) and an acyl glucuronide (AcMPAG). It is known that AcMPAG, which may be an immunotoxic metabolite, is deglucuronidated in human liver. AcMPAG deglucuronidation activity was detected in both human liver cytosol (HLC) and microsomes (HLM). By purification from HLC with column chromatographic purification steps, the enzyme responsible for AcMPAG deglucuronidationis identified as α/β hydrolase domain containing 10 (ABHD10). Recombinant ABHD10 expressed in Sf9 cells efficiently deglucuronidated AcMPAG with a K(m) value of 100.7 ± 10.2 μM, which was similar to those in HLM, HLC, and human liver homogenates (HLH). Immunoblot analysis revealed ABHD10 protein expression in both HLC and HLM. The AcMPAG deglucuronidation by recombinant ABHD10, HLC, and HLH were potently inhibited by AgNO(3), CdCl(2), CuCl(2), PMSF, bis-p-nitrophenylphosphate, and DTNB. The CL(int) value of AcMPAG formation from MPA, which was catalyzed by human UGT2B7, in HLH was increased by 1.8-fold in the presence of PMSF. Thus, human ABHD10 would affect the formation of AcMPAG, the immunotoxic metabolite.