Egfp p65

Manufactured by Addgene

The EGFP-p65 is a plasmid that expresses a fusion protein of enhanced green fluorescent protein (EGFP) and the p65 subunit of NF-kappa B. The EGFP-p65 plasmid can be used for monitoring the localization and dynamics of the p65 subunit in live cells.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Plentiekar2g2, by Addgene (1 mentions) Gw1 phred, by Addgene (1 mentions) Ptriex rhoa flare, by Addgene (1 mentions) Pmitotimer, by Addgene (1 mentions) Mcherry lifeact 7, by Addgene (1 mentions) Pgp cmv gcamp6f, by Addgene (1 mentions) Prk5 myc rhoa t19n, by Addgene (1 mentions) Lsm 780 microscope, by Zeiss (1 mentions)

2 protocols using egfp p65

Comprehensive RhoA Signaling Toolkit

Cited 1 time

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Raichu-RhoA (provided by M. Matsuda [27 (link)]), pTriEx-RhoA FLARE.sc Biosensor WT (RRID:Addgene_12150), pTriEx-RhoA FLARE.sc Biosensor Q63L (RRID:Addgene_12151), pTriEx-RhoA FLARE.sc Biosensor T19N (RRID:Addgene_12152), pRK5-myc-RhoA WT (RRID:Addgene_12962), pRK5-myc-RhoA Q63L (RRID:Addgene_12964), pRK5-myc-RhoA T19N (RRID:Addgene_12963), EGFP-p65 (RRID:Addgene_111190), GW1-pHRed (RRID:Addgene_31473), GW1CMV-Perceval (RRID:Addgene_21737), Laconic/pcDNA3.1 (+) (RRID:Addgene_118627), Pyronic /pcDNA3.1 (+) (RRID:Addgene_51308), pcDNA3.1 FLII12Pglu-700uDelta6 (RRID:Addgene_17866), Cyto-ABKAR (RRID:Addgene_61510), pLentiEKAR2G2 (RRID:Addgene_40178), Kras-Src FRET biosensor (RRID:Addgene_78302), pFRET-HSP33 cys (RRID:Addgene_16076), pGP-CMV-GCaMP6F (RRID:Addgene_40755), mCherry-Lifeact-7 (RRID:Addgene_54491), pMitoTimer (RRID:Addgene_52659), pCLBW cox8 EGFP mCherry (RRID:Addgene_78520).

Socodato R., Rodrigues-Santos A., Tedim-Moreira J., Almeida T.O., Canedo T., Portugal C.C, & Relvas J.B. (2023). RhoA balances microglial reactivity and survival during neuroinflammation. Cell Death & Disease, 14(10), 690.

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Visualizing p65 Nuclear Translocation

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WT and IKK2-KO VSMCs were seeded onto 35-mm glass-bottom imaging dishes (no. 1.5 cover glass). When cells reached approximately 70%–75% confluence, 2.5 μg of EGFP-p65 (Addgene, 111190) was transfected using Lipofectamine 2000 (Thermo Fisher Scientific). Twenty-four hours after transfection, cells were imaged in imaging buffer supplemented with 0.1% BSA and imaged with a Zeiss LSM 780 microscope with a C-Apochrome 40×/1.20 W Korr FCS M27 objective using the 488 nm wavelength laser in the presence of 1 μg/mL TNF-α at 1 frame every 5 seconds for 1 hour. Regions of interest were drawn over the nucleus and cytosol of cells and the nuclear fluorescent signal was divided by the cytosolic fluorescent signal to obtain the fluorescence ratio.

Miyazaki-Anzai S., Masuda M., Keenan A.L., Shiozaki Y., Miranda J.G, & Miyazaki M. (2024). Activation of the IKK2/NF-κB pathway in VSMCs inhibits calcified vascular stiffness in CKD. JCI Insight, 9(7), e174977.

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