Active Recombinant Full Length Mouse PIK3CA(E542K) and PIK3R1 Mutation Co-expressed Protein, N-His-tagged

• Cat.No.: PIK3CA-PIK3R1-139MFL
• Product Overview: Complex of N-terminal 6His-tagged recombinant murine p110α full length, containing the mutation E542K, and untagged, recombinant, murine p85α full length. Co-expressed by baculovirus in Sf21 insect cells. Purified using Ni2+/NTA-agarose. The E542K substitution is a somatic mutation in p110α that has been associated with tumours of the colon and brain. Combined in vitro and in vivo studies have shown that this mutation confers higher lipid kinase activity than wild type, and is able to induce oncogenic transformation.

Specification

• Description: Complex of N-terminal 6His-tagged recombinant, murine p110α full length, containing the mutation E542K, and untagged, recombinant, murine p85α full length. The E542K substitution is a somatic mutation in p110α that has been associated with tumors of the colon and brain. Combined in vitro and in vitro studies have shown that this mutation confers higher lipid kinase activity than wild type, and is able to induce oncogenic transformation.
• Source: Sf21
• Species: Mouse
• Tag: His
• Protein length: Full Length
• Form: 0.728mg/ml of enzyme in 50mM Tris/HCl pH7.5, 300mM NaCl, 0.1mM EGTA, 0.03% Brij-35, 270mM sucrose, 0.2mM PMSF, 1mM benzamidine, 0.1% 2-mercaptoethanol. Frozen solution.
• Bio-activity: 436U/mg, where one unit of PI 3-Kinase (p110α(E542K)/p85α)) activity is defined as 1nmol phosphatidylinositol 3,4,5-trisphosphate (PIP3) formed per minute at room temperature with a final ATP concentration of 100μM.
• Molecular Mass: p110α(E542K) MW = 129kDa, p85α MW = 84kDa
• AA Sequence: p110α(E542K) amino acid sequence: 1 MSYYHHHHHH DYDIPTTENL YFQGAMDPEF KGLRRQMPPR PSSGELWGIH LMPPRILVEC 61 LLPNGMIVTL ECLREATLVT IKHELFREAR KYPLHQLLQD ETSYIFVSVT QEAEREEFFD 121 ETRRLCDLRL FQPFLKVIEP VGNREEKILN REIGFVIGMP VCEFDMVKDP EVQDFRRNIL 181 NVCKEAVDLR DLNSPHSRAM YVYPPNVESS PELPKHIYNK LDKGQIIVVI WVIVSPNNDK 241 QKYTLKINHD CVPEQVIAEA IRKKTRSMLL SSEQLKLCVL EYQGKYILKV CGCDEYFLEK 301 YPLSQYKYIR SCIMLGRMPN LMLMAKESLY SQLPIDSFTM PSYSRRISTA TPYMNGETST 361 KSLWVINSAL RIKILCATYV NVNIRDIDKI YVRTGIYHGG EPLCDNVNTQ RVPCSNPRWN 421 EWLNYDIYIP DLPRAARLCL SICSVKGRKG AKEEHCPLAW GNINLFDYTD TLVSGKMALN 481 LWPVPHGLED LLNPIGVTGS NPNKETPCLE LEFDWFSSVV KFPDMSVIEE HANWSVSREA 541 GFSYSHTGLS NRLARDNELR ENDKEQLRAL CTRDPLSKIT EQEKDFLWSH RHYCVTIPEI 601 LPKLLLSVKW NSRDEVAQMY CLVKDWPPIK PEQAMELLDC NYPDPMVRSF AVRCLEKYLT 661 DDKLSQYLIQ LVQVLKYEQY LDNLLVRFLL KKALTNQRIG HFFFWHLKSE MHNKTVSQRF 721 GLLLESYCRA CGMYLKHLNR QVEAMEKLIN LTDILKQEKK DETQKVQMKF LVEQMRQPDF 781 MDALQGFLSP LNPAHQLGNL RLEECRIMSS AKRPLWLNWE NPDIMSELLF QNNEIIFKNG 841 DDLRQDMLTL QIIRIMENIW QNQGLDLRML PYGCLSIGDC VGLIEVVRNS HTIMQIQCKG 901 GLKGALQFNS HTLHQWLKDK NKGEIYDAAI DLFTRSCAGY CVATFILGIG DRHNSNIMVK 961 DDGQLFHIDF GHFLDHKKKK FGYKRERVPF VLTQDFLIVI SKGAQEYTKT REFERFQEMC 1021 YKAYLAIRQH ANLFINLFSM MLGSGMPELQ SFDDIAYIRK TLALDKTEQE ALEYFTKQMN 1081 DAHHGGWTTK MDWIFHTIKQ HALN
  p85α amino acid sequence:
  1 MSAEGYQYRA LYDYKKEREE DIDLHLGDIL TVNKGSLVAL GFSDGQEARP EDIGWLNGYN
  61 ETTGERGDFP GTYVEYIGRK RISPPTPKPR PPRPLPVAPG SSKTEADTEQ QALPLPDLAE
  121 QFAPPDVAPP LLIKLLEAIE KKGLECSTLY RTQSSSNPAE LRQLLDCDAA SVDLEMIDVH
  181 VLADAFKRYL ADLPNPVIPV AVYNEMMSLA QELQSPEDCI QLLKKLIRLP NIPHQCWLTL
  241 QYLLKHFFKL SQASSKNLLN ARVLSEIFSP VLFRFPAASS DNTEHLIKAI EILISTEWNE
  301 RQPAPALPPK PPKPTTVANN SMNNNMSLQD AEWYWGDISR EEVNEKLRDT ADGTFLVRDA
  361 STKMHGDYTL TLRKGGNNKL IKIFHRDGKY GFSDPLTFNS VVELINHYRN ESLAQYNPKL
  421 DVKLLYPVSK YQQDQVVKED NIEAVGKKLH EYNTQFQEKS REYDRLYEEY TRTSQEIQMK
  481 RTAIEAFNET IKIFEEQCQT QERYSKEYIE KFKREGNEKE IQRIMHNHDK LKSRISEIID
  541 SRRRLEEDLK KQAAEYREID KRMNSIKPDL IQLRKTRDQY LMWLTQKGVR QKKLNEWLGN
  601 ENTEDQYSLV EDDEDLPHHD EKTWNVGSSN RNKAENLLRG KRDGTFLVRE SSKQGCYACS
  661 VVVDGEVKHC VINKTATGYG FAEPYNLYSS LKELVLHYQH TSLVQHNDSL NVTLAYPVYA
  721 QQRR
  
• Purity: Purity (p110α(E542K) and p85α combined) 75.2% by SDS-PAGE and Coomassie blue staining.
• Applications: Drug Discovery & Development
• Storage: 6 months at -70 centigrade.
• Handling Advice: Rapidly thaw the vial under cold water and immediately place on ice. Aliquot unused material into prechilled micro-centrifuge tubes and immediately snap-freeze the vials in liquid nitrogen prior to re-storage at -70 centigrade.
• Use/Stability: On receipt of material store at -70 centigrade. Unopened reagent is stable for a minimum of 1 year from date of shipment when stored at recommended storage temperature. Avoid repeat freeze/thaw cycles. For maximum recovery of product, centrifuge original vial prior to removing the cap.
• References: Kang S. et al., PNAS, (2005);102: 802-807 and Zhao J.J. et al., PNAS, (2005);102:18443-18448.

Gene Information

• Gene Name: Pik3ca phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha [ Mus musculus (house mouse) ] and Pik3r1 phosphoinositide-3-kinase regulatory subunit 1 [ Mus musculus (house mouse) ]
• Official Symbol: PIK3CA and PIK3R1
• Synonyms: PIK3CA and PIK3R1; phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha and phosphoinositide-3-kinase regulatory subunit 1; p110; caPI3K; p110alpha; 6330412C24Rik and PI3K; p50alpha; p55alpha; p85alpha
• Gene ID: 18706 and 18708
• mRNA Refseq: NM_008839.3 and NM_001024955.2
• Protein Refseq: NP_032865.2 and NP_001020126.1
• UniProt ID: P42337 and P26450