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Human AMPK (G1/B1/A2) Heterotrimer Insect Cell Lysate (WB positive control)

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    Human PRKAA1 Transfected / Overexpression Cell Lysate Product Information
    Expressed Host:Baculovirus-Insect cells
    Product Description:Baculovirus-Insect Cell lysate that Human AMPK (G1/B1/A2) Heterotrimer transfected / overexpressed for Western blot (WB) positive control. The whole cell lysate is provided in 1X Sample Buffer (1X modified RIPA buffer+1X SDS loading buffer).
    Sequence information:A DNA sequence encoding the human PRKAG1 (P54619) (Met 1-Pro 331), constructed the plasmid 1; A DNA sequence encoding the human PRKAB1 (Q9Y478) (Met 1-Ile 270), constructed the plasmid 2; A DNA sequence encoding the human PRKAA2 (P54646) (Met 1-Arg 552) was fused with the N-terminal polyhistidine-tagged GST tag at the N-terminus, constructed the plasmid 3. The three plasmids were co-expressed and the heterotrimer was purified.
    Predicted N Terminal:Met & Met & His
    Molecule Mass:The recombinant heterotrimer of human AMPK (PRKAG1 / PRKAB1 / PRKAA2) has a calculated molecular mass of 158 (38+30+90) KDa. The apparent molecular mass is approximately 40, 42 & 95 KDa respectively in SDS-PAGE under reducing conditions.
    Species:Human
    Human PRKAA1 Transfected / Overexpression Cell Lysate Usage Guide
    Preparation Method:Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
    Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
    Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue after protein purification.
    Stability:Samples are stable for up to twelve months from date of receipt at -70℃
    Recommend Usage:1.  Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2.  Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
    Storage Buffer:1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
    Storage Instruction:Store at 4℃. After re-dissolution, aliquot and store at -80℃.
    Application notes:Western blot (WB): Use at an assay dependent dilution.
    Other Applications: Not tested.
    Optimal dilutions/concentrations should be determined by the end user.
    All information of our products is subject to change without notice. Please refer to COA enclosed in shipped package for the newest information.
    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"