IDE (NM_004969) Human Tagged ORF Clone

CAT#: RC220700

IDE (Myc-DDK-tagged)-Human insulin-degrading enzyme (IDE), transcript variant 1

Product Images

Circular map for RC220700

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  Western blot validation of overexpression lysate (Cat# LY401545) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC220700 using transfection reagent MegaTran 2.0 (Cat# TT210002).

• 

  Coomassie blue staining of purified IDE protein (Cat# TP320700). The protein was produced from HEK293T cells transfected with IDE cDNA clone (Cat# RC220700) using MegaTran 2.0 (Cat# TT210002).

• 

  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY IDE (Cat# RC220700, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-IDE (Cat# TA811776)(1:500).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: IDE
• Synonyms: INSULYSIN
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC220700 representing NM_004969
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGCGGTACCGGCTAGCGTGGCTTCTGCACCCCGCACTGCCCAGCACCTTCCGCTCAGTCCTCGGCGCCC
  GCCTGCCGCCTCCGGAGCGCCTGTGTGGTTTCCAAAAAAAGACTTACAGCAAAATGAATAATCCAGCCAT
  CAAGAGAATAGGAAATCACATTACCAAGTCTCCTGAAGACAAGCGAGAATATCGAGGGCTAGAGCTGGCC
  AATGGTATCAAAGTACTTCTTATGAGTGATCCCACCACGGATAAGTCATCAGCAGCACTTGATGTGCACA
  TAGGTTCATTGTCGGATCCTCCAAATATTGCTGGCTTAAGTCATTTTTGTGAACATATGCTTTTTTTGGG
  AACAAAGAAATACCCTAAAGAAAATGAATACAGCCAGTTTCTCAGTGAGCATGCAGGAAGTTCAAATGCC
  TTTACTAGTGGAGAGCATACCAATTACTATTTTGATGTTTCTCATGAACACCTAGAAGGTGCCCTAGACA
  GGTTTGCACAGTTTTTTCTGTGCCCCTTGTTCGATGAAAGTTGCAAAGACAGAGAGGTGAATGCAGTTGA
  TTCAGAACATGAGAAGAATGTGATGAATGATGCCTGGAGACTCTTTCAATTGGAAAAAGCTACAGGGAAT
  CCTAAACACCCCTTCAGTAAATTTGGGACAGGTAACAAATATACTCTGGAGACTAGACCAAACCAAGAAG
  GCATTGATGTAAGACAAGAGCTACTGAAATTCCATTCTGCTTACTATTCATCCAACTTAATGGCTGTTTG
  TGTTTTAGGTCGAGAATCTTTAGATGACTTGACTAATCTGGTGGTAAAGTTATTTTCTGAAGTAGAGAAC
  AAAAATGTTCCATTGCCAGAATTTCCTGAACACCCTTTCCAAGAAGAACATCTTAAACAACTTTACAAAA
  TAGTACCCATTAAAGATATTAGGAATCTCTATGTGACATTTCCCATACCTGACCTTCAGAAATACTACAA
  ATCAAATCCTGGTCATTATCTTGGTCATCTCATTGGGCATGAAGGTCCTGGAAGTCTGTTATCAGAACTT
  AAGTCAAAGGGCTGGGTTAATACTCTTGTTGGTGGGCAGAAGGAAGGAGCCCGAGGTTTTATGTTTTTTA
  TCATTAATGTGGACTTGACCGAGGAAGGATTATTACATGTTGAAGATATAATTTTGCACATGTTTCAATA
  CATTCAGAAGTTACGTGCAGAAGGACCTCAAGAATGGGTTTTCCAAGAGTGCAAGGACTTGAATGCTGTT
  GCTTTTAGGTTTAAAGACAAAGAGAGGCCACGGGGCTATACATCTAAGATTGCAGGAATATTGCATTATT
  ATCCCCTAGAAGAGGTGCTCACAGCGGAATATTTACTGGAAGAATTTAGACCTGACTTAATAGAGATGGT
  TCTCGATAAACTCAGACCAGAAAATGTCCGGGTTGCCATAGTTTCTAAATCTTTTGAAGGAAAAACTGAT
  CGCACAGAAGAGTGGTATGGAACCCAGTACAAACAAGAAGCTATACCGGATGAAGTCATCAAGAAATGGC
  AAAATGCTGACCTGAATGGGAAATTTAAACTTCCTACAAAGAATGAATTTATTCCTACGAATTTTGAGAT
  TTTACCGTTAGAAAAAGAGGCGACACCATACCCTGCTCTTATTAAGGATACAGTCATGAGCAAACTTTGG
  TTCAAACAAGATGATAAGAAAAAAAAGCCGAAGGCTTGTCTCAACTTTGAATTTTTCAGCCCATTTGCTT
  ATGTGGACCCCTTGCACTGTAACATGGCCTATTTGTACCTTGAGCTCCTCAAAGACTCACTCAACGAGTA
  TGCATATGCAGCAGAGCTAGCAGGCTTGAGCTATGATCTCCAAAATACCATCTATGGGATGTATCTTTCA
  GTGAAAGGTTACAATGACAAGCAGCCAATTTTACTAAAGAAGATTATTGAGAAAATGGCTACCTTTGAGA
  TTGATGAAAAAAGATTTGAAATTATCAAAGAAGCATATATGCGATCTCTTAACAATTTCCGGGCTGAACA
  GCCTCACCAGCATGCCATGTACTACCTCCGCTTGCTGATGACTGAAGTGGCCTGGACTAAAGATGAGTTA
  AAAGAAGCTCTGGATGATGTAACCCTTCCTCGCCTTAAGGCCTTCATACCTCAGCTCCTGTCACGGCTGC
  ACATTGAAGCCCTTCTCCATGGAAACATAACAAAGCAGGCTGCATTAGGAATTATGCAGATGGTTGAAGA
  CACCCTCATTGAACATGCTCATACCAAACCTCTCCTTCCAAGTCAGCTGGTTCGGTATAGAGAAGTTCAG
  CTCCCTGACAGAGGATGGTTTGTTTATCAGCAGAGAAATGAAGTTCACAATAACTGTGGCATCGAGATAT
  ACTACCAAACAGACATGCAAAGCACCTCAGAGAATATGTTTCTGGAGCTCTTCTGTCAGATTATCTCGGA
  ACCTTGCTTCAACACCCTGCGCACCAAGGAGCAGTTGGGCTATATCGTCTTCAGCGGGCCACGTCGAGCT
  AATGGCATACAGAGCTTGAGATTCATCATCCAGTCAGAAAAGCCACCTCACTACCTAGAAAGCAGAGTGG
  AAGCTTTCTTAATTACCATGGAAAAGTCCATAGAGGACATGACAGAAGAGGCCTTCCAAAAACACATTCA
  GGCATTAGCAATTCGTCGACTAGACAAACCAAAGAAGCTATCTGCTGAGTGTGCTAAATACTGGGGAGAA
  ATCATCTCCCAGCAATATAATTTTGACAGAGATAACACTGAGGTTGCATATTTAAAGACACTTACCAAGG
  AAGATATCATCAAATTCTACAAGGAAATGTTGGCAGTAGATGCTCCAAGGAGACATAAGGTATCCGTCCA
  TGTTCTTGCCAGGGAAATGGATTCTTGTCCTGTTGTTGGAGAGTTCCCATGTCAAAATGACATAAATTTG
  TCACAAGCACCAGCCTTGCCACAACCTGAAGTGATTCAGAACATGACCGAATTCAAGCGTGGTCTGCCAC
  TGTTTCCCCTTGTGAAACCACATATTAACTTCATGGCTGCAAAACTC
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC220700 representing NM_004969
  Red=Cloning site Green=Tags(s)
  
  MRYRLAWLLHPALPSTFRSVLGARLPPPERLCGFQKKTYSKMNNPAIKRIGNHITKSPEDKREYRGLELA
  NGIKVLLMSDPTTDKSSAALDVHIGSLSDPPNIAGLSHFCEHMLFLGTKKYPKENEYSQFLSEHAGSSNA
  FTSGEHTNYYFDVSHEHLEGALDRFAQFFLCPLFDESCKDREVNAVDSEHEKNVMNDAWRLFQLEKATGN
  PKHPFSKFGTGNKYTLETRPNQEGIDVRQELLKFHSAYYSSNLMAVCVLGRESLDDLTNLVVKLFSEVEN
  KNVPLPEFPEHPFQEEHLKQLYKIVPIKDIRNLYVTFPIPDLQKYYKSNPGHYLGHLIGHEGPGSLLSEL
  KSKGWVNTLVGGQKEGARGFMFFIINVDLTEEGLLHVEDIILHMFQYIQKLRAEGPQEWVFQECKDLNAV
  AFRFKDKERPRGYTSKIAGILHYYPLEEVLTAEYLLEEFRPDLIEMVLDKLRPENVRVAIVSKSFEGKTD
  RTEEWYGTQYKQEAIPDEVIKKWQNADLNGKFKLPTKNEFIPTNFEILPLEKEATPYPALIKDTVMSKLW
  FKQDDKKKKPKACLNFEFFSPFAYVDPLHCNMAYLYLELLKDSLNEYAYAAELAGLSYDLQNTIYGMYLS
  VKGYNDKQPILLKKIIEKMATFEIDEKRFEIIKEAYMRSLNNFRAEQPHQHAMYYLRLLMTEVAWTKDEL
  KEALDDVTLPRLKAFIPQLLSRLHIEALLHGNITKQAALGIMQMVEDTLIEHAHTKPLLPSQLVRYREVQ
  LPDRGWFVYQQRNEVHNNCGIEIYYQTDMQSTSENMFLELFCQIISEPCFNTLRTKEQLGYIVFSGPRRA
  NGIQSLRFIIQSEKPPHYLESRVEAFLITMEKSIEDMTEEAFQKHIQALAIRRLDKPKKLSAECAKYWGE
  IISQQYNFDRDNTEVAYLKTLTKEDIIKFYKEMLAVDAPRRHKVSVHVLAREMDSCPVVGEFPCQNDINL
  SQAPALPQPEVIQNMTEFKRGLPLFPLVKPHINFMAAKL
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_004969
• ORF Size: 3057 bp
• OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info
• OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene.
• Product Components: The ORF clone is ion-exchange column purified and shipped in a 2D barcoded Matrix tube containing 10ug of transfection-ready, dried plasmid DNA (reconstitute with 100 ul of water).
• Reconstitution: 1. Centrifuge at 5,000xg for 5min.
  2. Carefully open the tube and add 100ul of sterile water to dissolve the DNA.
  3. Close the tube and incubate for 10 minutes at room temperature.
  4. Briefly vortex the tube and then do a quick spin (less than 5000xg) to concentrate the liquid at the bottom.
  5. Store the suspended plasmid at -20°C. The DNA is stable for at least one year from date of shipping when stored at -20°C.

Reference Data

• RefSeq: NM_004969.1, NP_004960.1
• RefSeq Size: 3279 bp
• RefSeq ORF: 3060 bp
• Locus ID: 3416
• UniProt ID: P14735
• Cytogenetics: 10q23.33
• Domains: Peptidase_M16, Peptidase_M16_C
• Protein Families: Druggable Genome, Protease
• Protein Pathways: Alzheimer's disease
• MW: 117.8 kDa
• Gene Summary: This gene encodes a zinc metallopeptidase that degrades intracellular insulin, and thereby terminates insulins activity, as well as participating in intercellular peptide signalling by degrading diverse peptides such as glucagon, amylin, bradykinin, and kallidin. The preferential affinity of this enzyme for insulin results in insulin-mediated inhibition of the degradation of other peptides such as beta-amyloid. Deficiencies in this protein's function are associated with Alzheimer's disease and type 2 diabetes mellitus but mutations in this gene have not been shown to be causitive for these diseases. This protein localizes primarily to the cytoplasm but in some cell types localizes to the extracellular space, cell membrane, peroxisome, and mitochondrion. Alternative splicing results in multiple transcript variants encoding distinct isoforms. Additional transcript variants have been described but have not been experimentally verified.[provided by RefSeq, Sep 2009]