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

• Cat.No.: PIK3CA-RELA-131HFL
• Product Overview: Complex of N-terminal 6Histagged, recombinant, full-length human p110α and untagged, recombinant, full-length human p65α. Coexpressed by baculovirus in Sf21 insect cells and purified using Ni2+/NTA-agarose. p65α is an oncogenic form of p85α 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: Human
• Tag: His
• Protein length: Full Length
• Form: 1.927mg/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: 596U/mg, where one unit of PI 3-Kinase alpha (p110α/p65α (h)) activity is defined as 1nmol phosphatidylinositol 3,4,5- trisphosphate (PIP3) formed per minute at room temperature with a final ATP concentration of 100M.
• Molecular Mass: p110α MW = 125.3kDa, p65α MW = 66.0kDa
• AA Sequence: p110α amino acid sequence: 1 MHHHHHHMPP RPSSGELWGI HLMPPRILVE CLLPNGMIVT LECLREATLI TIKHELFKEA 61 RKYPLHQLLQ DESSYIFVSV TQEAEREEFF DETRRLCDLR LFQPFLKVIE PVGNREEKIL 121 NREIGFAIGM PVCEFDMVKD PEVQDFRRNI LNVCKEAVDL RDLNSPHSRA MYVYPPNVES 181 SPELPKHIYN KLDKGQIIVV IWVIVSPNND KQKYTLKINH DCVPEQVIAE AIRKKTRSML 241 LSSEQLKLCV LEYQGKYILK VCGCDEYFLE KYPLSQYKYI RSCIMLGRMP NLMLMAKESL 301 YSQLPMDCFT MPSYSRRIST ATPYMNGETS TKSLWVINSA LRIKILCATY VNVNIRDIDK 361 IYVRTGIYHG GEPLCDNVNT QRVPCSNPRW NEWLNYDIYI PDLPRAARLC LSICSVKGRK 421 GAKEEHCPLA WGNINLFDYT DTLVSGKMAL NLWPVPHGLE DLLNPIGVTG SNPNKETPCL 481 ELEFDWFSSV VKFPDMSVIE EHANWSVSRE AGFSYSHAGL SNRLARDNEL RENDKEQLKA 541 ISTRDPLSEI TEQEKDFLWS HRHYCVTIPE ILPKLLLSVK WNSRDEVAQM YCLVKDWPPI 601 KPEQAMELLD CNYPDPMVRG FAVRCLEKYL TDDKLSQYLI QLVQVLKYEQ YLDNLLVRFL 661 LKKALTNQRI GHFFFWHLKS EMHNKTVSQR FGLLLESYCR ACGMYLKHLN RQVEAMEKLI 721 NLTDILKQEK KDETQKVQMK FLVEQMRRPD FMDALQGFLS PLNPAHQLGN LRLEECRIMS 781 SAKRPLWLNW ENPDIMSELL FQNNEIIFKN GDDLRQDMLT LQIIRIMENI WQNQGLDLRM 841 LPYGCLSIGD CVGLIEVVRN SHTIMQIQCK GGLKGALQFN SHTLHQWLKD KNKGEIYDAA 901 IDLFTRSCAG YCVATFILGI GDRHNSNIMV KDDGQLFHID FGHFLDHKKK KFGYKRERVP 961 FVLTQDFLIV ISKGAQECTK TREFERFQEM CYKAYLAIRQ HANLFINLFS MMLGSGMPEL 1021 QSFDDIAYIR KTLALDKTEQ EALEYFMKQM NDAHHGGWTT KMDWIFHTIK QHALN
  p65α amino acid sequence:
  1 MSAEGYQYRA LYDYKKEREE DIDLHLGDIL TVNKGSLVAL GFSDGQEARP EEIGWLNGYN
  61 ETTGERGDFP GTYVEYIGRK KISPPTPKPR PPRPLPVAPG SSKTEADVEQ QALTLPDLAE
  121 QFAPPDIAPP LLIKLVEAIE KKGLECSTLY RTQSSSNLAE LRQLLDCDTP SVDLEMIDVH
  181 VLADAFKRYL LDLPNPVIPA AVYSEMISLA PEVQSSEEYI QLLKKLIRSP SIPHQYWLTL
  241 QYLLKHFFKL SQTSSKNLLN ARVLSEIFSP MLFRFSAASS DNTENLIKVI EILISTEWNE
  301 RQPAPALPPK PPKPTTVANN GMNNNMSLQD AEWYWGDISR EEVNEKLRDT ADGTFLVRDA
  361 STKMHGDYTL TLRKGGNNKL IKIFHRDGKY GFSDPLTFSS VVELINHYRN ESLAQYNPKL
  421 DVKLLYPVSK YQQDQVVKED NIEAVGKKLH EYNTQFQEKS REYDRLYEEY TRTSQEIQMK
  481 RTAIEAFNET IKIFEEQCQT QERYSKEYIE KFKREGNEKE IQRIMHNYDK LKSRISEIID
  541 SRRRLEEDLK KQAAEYREID KRMNSIKPDL I
  
• Purity: Purity (p110α & p65α combined) 86% 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: 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 [ Homo sapiens (human) ] and RELA RELA proto-oncogene, NF-kB subunit [ Homo sapiens (human) ]
• 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; MCM; CCM4; CWS5; MCAP; PI3K; CLAPO; CLOVE; MCMTC; PI3K-alpha; p110-alpha and p65; CMCU; NFKB3; AIF3BL3
• Gene ID: 5290 and 5970
• mRNA Refseq: NM_006218.4 and NM_001145138.2
• Protein Refseq: NP_006209.2 and NP_001138610.1
• MIM: 171834 and 164014
• UniProt ID: P42336 and Q04206