Pcdna3 flag ha akt1

Manufactured by Addgene

Sourced in United States

The pcDNA3-Flag-HA-Akt1 is a plasmid vector that expresses the Akt1 protein with an N-terminal Flag and HA tag. Akt1 is a serine/threonine-protein kinase that plays a key role in multiple cellular processes including cell proliferation, survival, and metabolism. The plasmid is commonly used in research to study the function and regulation of Akt1.

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Lab products found in correlation

Pcdna3 myr ha akt1, by Addgene (1 mentions) Phiv irfp720 e2a luc, by Addgene (1 mentions) Plncx myr ha akt1, by Addgene (1 mentions) Quickchange lightning site directed mutagenesis kit, by Agilent Technologies (1 mentions)

Spelling variants of this product

PcDNA3.1 (119 citations) FLAG-HA-pcDNA3.1 (25 citations) PcDNA3-HA-AKT1 (5 citations) PcDNA3-HA (3 citations) Flag-pcDNA3.1 (2 citations)

2 protocols using pcdna3 flag ha akt1

Plasmid Expression Constructs for FOXO Signaling

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Expressing plasmids, including pcDNA3-Flag-FKHR(FOXO1)-Delta DB, pcDNA3-flag FKHRL1 (FOXO3a), pcDNA3-Flag-HA, pcDNA3-Flag-HA-Akt1, pcDNA3-Myr-HA-Akt1, and pcDNA3-T7-Akt1-T308A-S473A [34 (link)] were purchased from Addgene (Cambridge, MA, USA). The HBP1 expressing plasmid pEF-BOS-HA-HBP1, and FOXO1 expressing plasmids pcDNA3-A3-FOXO1-Flag pcDNA3-WT-FOXO1-Flag and pcDNA3-H215R-FOXO1-Flag were kindly provided by Dr. Amy S. Yee and Dr. Brian Schaffhausen (Tufts University, USA), respectively.

Chan C.Y., Huang S.Y., Sheu J.J., Roth M.M., Chou I.T., Lien C.H., Lee M.F, & Huang C.Y. (2017). Transcription factor HBP1 is a direct anti-cancer target of transcription factor FOXO1 in invasive oral cancer. Oncotarget, 8(9), 14537-14548.

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Cloning and Mutagenesis of AKT1 and PRMT5

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Human pCDNA3/Flag-HA-AKT1, Plncx/myr-HA-AKT1, pCDNA6A/EGFR, and pHIV-iRFP720-E2A-Luc were purchased from Addgene. pCDNA3.1/Flag-PRMT5 wild type and dead enzyme mutant were as described previously^{55 (link)}. pCDNA3/Flag-HA-AKT1-R15K mutant was generated by site mutagenesis using QuickChange Lightning Site-Directed Mutagenesis Kit (Agilent, cat#210518) following the manufacturer’s protocol. Primers introducing mutation sites were designed by Agilent PrimerDesign Program on the Agilent website, AKT1-R15K_forward 5′-GATGTACTCCCCTTTTTTGTGCAGCCAACCCTCCTTCACA-3′, AKT1-R15K_reverse 5′-TGTGAAGGAGGGTTGGCTGCACAAAAAAGGGGAGTACATC-3′. pLKO-Tet-ON inducible shPRMT5 constructs were generous gifts from Dr. William R. Sellers (Broad Institute). siRNA targeting PRMT9 was purchased from Thermo Fisher Scientific Inc. (cat#4392420).

Huang L., Zhang X.O., Rozen E.J., Sun X., Sallis B., Verdejo-Torres O., Wigglesworth K., Moon D., Huang T., Cavaretta J.P., Wang G., Zhang L., Shohet J.M., Lee M.M, & Wu Q. (2022). PRMT5 activates AKT via methylation to promote tumor metastasis. Nature Communications, 13, 3955.

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