Active Recombinant Full Length Mouse PIK3CA and RELA Co-expressed Protein, N-His-tagged

• Cat.No.: PIK3CA-RELA-132MFL
• Product Overview: Complex of N-terminal 6Histagged recombinant mouse p110α full length and untagged, recombinant, mouse p65α full length. Coexpressed by baculovirus in Sf21 insect cells. Purified using Ni2+/NTA-agarose. p65α is an oncogenic form of p65α that binds but does not inhibit p110, leading to constitutive PI3K activity

Specification

• Description: PI3 Kinase plays a key role by recruiting PH domain-containing proteins to the membrane, including AKT1 and PDPK1, activating signaling cascades involved in cell growth, survival, proliferation, motility and morphology. It also participates in cellular signaling in response to various growth factors and involved in the activation of AKT1 upon stimulation by receptor tyrosine kinases ligands such as EGF, insulin, IGF1, VEGFA and PDGF. PI3K regulates invadopodia formation through the PDPK1-AKT1 pathway, and participates in cardiomyogenesis in embryonic stem cells through a AKT1 pathway. In addition to its lipid kinase activity, it displays a serine-protein kinase activity that results in the autophosphorylation of the p85alpha regulatory subunit as well as phosphorylation of other proteins such as 4EBP1, H-Ras, the IL-3 beta c receptor and possibly others.
• Source: Sf21
• Species: Mouse
• Tag: His
• Protein length: Full Length
• Form: 1.165mg/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: 611U/mg, where one unit of PI 3-Kinase (p110α/p65α) 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 MW = 129kDa, p65α MW = 65.9kDa
• AA Sequence: p110α 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 CTRDPLSEIT 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
  p65α 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 I
• Purity: Purity (p110α and p65α combined) 81.5% by SDSPAGE 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: Shekar, S.C. et al., J. Biol. Chem., (2005);280: 27850- 27855 and Jimenez, C. et al., EMBO J.,(1998);17:743- 753.

Gene Information

• Gene Name: Pik3ca phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha [ Mus musculus (house mouse) ] and Rela v-rel reticuloendotheliosis viral oncogene homolog A (avian) [ Mus musculus (house mouse) ]
• Official Symbol: PIK3CA and Rela
• Synonyms: PIK3CA and Rela; phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha and RELA proto-oncogene, NF-kB subunit; p110; caPI3K; p110alpha; 6330412C24Rik and p65; p65 NFkB; p65 NF-kappa B
• Gene ID: 18706 and 19697
• mRNA Refseq: NM_008839.3 and NM_009045.5
• Protein Refseq: NP_032865.2 and NP_033071.1
• UniProt ID: P42337 and Q548Y4