Pcdna3 flag lkb1

Manufactured by Addgene

Sourced in Spain

pCDNA3-FLAG-LKB1 is a plasmid vector that expresses the LKB1 (Liver Kinase B1) protein with a FLAG tag. LKB1 is a serine/threonine protein kinase that plays a role in the regulation of cellular energy metabolism and cell growth.

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Lab products found in correlation

Geneticin, by Thermo Fisher Scientific (1 mentions) Anti β actin, by Merck Group (1 mentions) Anti gapdh, by Merck Group (1 mentions) Hygromycin, by Merck Group (1 mentions) Inpp4b, by Cell Signaling Technology (1 mentions) Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (1 mentions) Jetprime reagent, by Polyplus Transfection (1 mentions) Pcdna3 lacz, by Thermo Fisher Scientific (1 mentions) Gapdh, by Merck Group (1 mentions) Sodium palmitate, by Merck Group (1 mentions) Fa free bsa, by Merck Group (1 mentions) Sicd36, by Thermo Fisher Scientific (1 mentions) Anti fyn, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

PcDNA3.1 (119 citations) Flag-pcDNA3.1 (2 citations)

4 protocols using pcdna3 flag lkb1

Overexpression of LKB1 in A549 cells

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A549 cells were transfected with pcDNA3-FLAG-LKB1 (plasmid 8590 [29 (link)], Addgene, Cambridge MA) to overexpress LKB1 or with pcDNA3.1 as vector control using GeneJuice transfection reagent (Novagen, Etobicoke, ON). Transfected cells were seeded into multiple tissue culture dishes and after 48 hours, actively dividing cells were treated with 500 μg/ml Geneticin (Invitrogen, Burlington, ON) to facilitate selection of plasmid-expressing cells. Cells in each tissue culture dish remaining after selection were pooled to create a number of pooled clones of both LKB1 expressing and non-expressing A549 cells. Confirmation of LKB1 expression in FLAG-LKB1 transfected cells and continued lack of LKB1 expression in control cells was confirmed by western blotting for LKB1.

Howe G.A., Xiao B., Zhao H., Al-Zahrani K.N., Hasim M.S., Villeneuve J., Sekhon H.S., Goss G.D., Sabourin L.A., Dimitroulakos J, & Addison C.L. (2016). Focal Adhesion Kinase Inhibitors in Combination with Erlotinib Demonstrate Enhanced Anti-Tumor Activity in Non-Small Cell Lung Cancer. PLoS ONE, 11(3), e0150567.

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Antibody-based Evaluation of AMPK Signaling

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Antibodies against AMPKα, phospho-AMPKα (Thr¹⁷²), LKB1 and INPP4B were purchased from Cell Signaling (Beverly, MA). Anti-LKB1 for immunohistochemistry, anti-β-actin, anti-GAPDH, G418 and hygromycin were obtained from Sigma-Aldrich (St. Louis, MO). The plasmid pc-DNA3-flag-LKB1 was bought from Addgene (Cambridge, MA). Lipofectamine 2000 transfection reagent was purchased from Invitrogen (Carlsbad, CA).

Zhang X., Chen H., Wang X., Zhao W, & Chen J.J. (2014). Expression and transcriptional profiling of the LKB1 tumor suppressor in cervical cancer cells. Gynecologic oncology, 134(2), 372-378.

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Hepatocyte Transfection for LKB1 and Akt Studies

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Phb1-KO hepatocytes were transfected with 2 μg of pcDNA3-FLAG-LKB1 (Addgene), pLNCX1-HA-Akt (kindly provided by Dr. Carracedo, CIC bioGUNE, Spain) or cysteine protease (NEDP1-V5) plasmids using jetPRIME^TM reagent (Polyplus). Two μg of pcDNA3-LacZ (Invitrogen) and pLNCX1 plasmids (provided by Dr. Carracedo) were used as negative controls of LKB1 and Akt, respectively.

Barbier-Torres L., Delgado T.C., García-Rodríguez J.L., Zubiete-Franco I., Fernández-Ramos D., Buqué X., Cano A., Juan V.G., Fernández-Domínguez I., Lopitz-Otsoa F., Fernández-Tussy P., Boix L., Bruix J., Villa E., Castro A., Lu S.C., Aspichueta P., Xirodimas D., Varela-Rey M., Mato J.M., Beraza N, & Martínez-Chantar M.L. (2014). Stabilization of LKB1 and Akt by neddylation regulates energy metabolism in liver cancer. Oncotarget, 6(4), 2509-2523.

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Modulation of CD36 and LKB1 in Oxidized LDL Signaling

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FA-free BSA, sodium palmitate, and other chemicals were from Sigma-Aldrich (St. Louis, MO); and human highly oxidized LDL (oxLDL) was from Kalen Biomedical (Montgomery Village, MD). All phospho-antibodies were from Cell Signaling Technology (Danvers, MA), anti-Fyn was from Santa Cruz Biotechnology (Santa Cruz, CA), GAPDH was from Sigma-Aldrich, and anti-LKB1 was from Millipore (Billerica, MA). IRDye800 anti-rabbit or anti-goat IgG and IRDye700 anti-mouse IgG were from Rockland (Gilbertsville, PA). Anti-human and anti-mouse CD36 were from R&D Systems (Minneapolis, MN), rat monoclonal anti-mouse CD36 was from AbD Serotec (Raleigh, NC), and mouse monoclonal anti-human CD36 was from Abcam (Cambridge, MA). Alexa Fluor IgGs (594 anti-rabbit, 488 goat anti-mouse, and 488 donkey anti-goat) were from Life Technologies. Mouse CD36 small interfering RNAs (siRNA) (siCD36–1: 5′-AAACCCAGATGACGTGGCAAA-3′; siCD36–2: 5′-AACGACATGATTAATGGCACACCTGTCTC–3′) were from Life Technologies; and pcDNA3-FLAG-LKB1 (catalog #8591), pRK5 c-Fyn (catalog #16032), and pRK5 DN-Fyn (catalog #16033) were from Addgene.

Samovski D., Sun J., Pietka T., Gross R.W., Eckel R.H., Su X., Stahl P.D, & Abumrad N.A. (2014). Regulation of AMPK Activation by CD36 Links Fatty Acid Uptake to β-Oxidation. Diabetes, 64(2), 353-359.

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