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fluorescent-protein-34px-icon.jpg Fluorescent Protein Guide: Subcellular Localization


Background

The subcellular localization of a protein is often tied to its function, so it's important to determine where your protein of interest resides. There are a variety of methods for determining subcellular localization. One of the most common is to use fluorescence microscopy to compare the localization of your protein of interest with known markers.

Addgene has assembled a collection of fluorescent fusion proteins that are targeted to various organelles or subcellular structures. You should fuse your protein of interest with a fluorescent protein that has a different emission wavelength. For example, if the marker protein is fused to GFP, you may want to fuse your protein to a red fluorescent protein (see this fusion protein guide to find the appropriate backbone). Then, by examining the overlap between your fusion protein and the marker fusion protein using fluorescence microscopy, you can assess whether your protein is targeted to the same structures as these well-characterized proteins.


General Structures

Browse or search the table below to find markers of general cellular organelles and structures.

Plasmid Structure Targeting Gene/Signal Fluorescent Protein PI
H2B-mCherry Chromatin H2B mCherry Benezra
H2B-GFP Chromatin H2B GFP Wahl
pCAG-mGFP Membrane palmitoylation sequence from GAP43 EGFP Cepko
pPalmitoyl-mTurquoise2 Membrane palmitoylation sequence from p63 mTurquoise2 Gadella
pmTurquoise2-Mito Mitochondria COX8A(1-29) mTurquoise2 Gadella
pEBFP2-Nuc Nucleus NLS EBFP2 Campbell
pmTurquoise2-H2A Nucleus H2A mTurquoise2 Gadella
CAV1-mCherry Caveolae Cav1 mCherry Helenius
pABCb10-GFP Mitochondria ABCb10 GFP Shirihai
mCherry-Mito-7 Mitochondria Mitochondrialtargeting sequence(COX8A) mCherry* Davidson
mito-BFP Mitochondria mitochondrial targeting signal (COX4) TagBFP Voeltz
pmTurquoise2-NES Non-nucleus Nuclear Export Sequence mTurquoise2 Gadella
pQC NLS mCherry IX Nucleus NLS mCherry Cepko
pmTurquoise2-Peroxi Peroxisomes Peroxisomal Targeting Sequence mTurquoise2 Gadella
GFP-Mff Mitochondria-Outer Membrane Mff acGFP1 Voeltz
mCherry-LaminB1-10 Nuclear Envelope LaminB1 mCherry* Davidson
mCherry-LaminA-C-18 Nuclear Envelope LaminA1 mCherry* Davidson
pEGFP-C1-Fibrillarin Nucleolus Fibrillarin EGFP Huang
pmKalama1-Nuc Nucleus NLS mKamala1 Campbell
pQC NLS YFP IX Nucleus NLS YFP Cepko
pcDNA3.1-kappa-myc-dL5-2xG4S-TMst Cell surface (mammalian) PDGFR-derived dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez
pH2B-miRFP703 Chromatin H2B miRFP703 Verkhusha
pcDNA3.1-KozATG-dL5-2XG4S-mCer3 Cytosol, nucleoplasm None dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez
cfSGFP2-N Extracellular milieu cfSGFP2-N EGFP Wada
Lck-GFP Membrane palmitoylation sequence from Lck EGFP Green
pcDNA3.1-COXIV-COX8-dL5-2XG4S-mCer3 Mitochondria COX IV-derived (1-22 aa) import sequence and COX VIII signal peptide dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez
pMito-miRFP703 Mitochondria COX8A miRFP703 Verkhusha
mCherry-TOMM20-N-10 Mitochondria-Outer Membrane TOMM20 mCherry* Davidson
pmScarlet-i_NES_C1 Non-nucleus Nuclear Export Sequence mScarlet-I Gadella
pmScarlet-H_NES_C1 Non-nucleus Nuclear Export Sequence mScarlet-H Gadella
pmScarlet_NES_C1 Non-nucleus Nuclear Export Sequence mScarlet Gadella
pcDNA3.1-NLS-myc-dL5-2xG4S-mCer3 Nucleus NLS (from Mak16p protein) dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez
pmScarlet-i_H2A_C1 Nucleus H2A mScarlet-I Gadella
pmScarlet-H_H2A_C1 Nucleus H2A mScarlet-H Gadella
pmScarlet_H2A_C1 Nucleus H2A mScarlet Gadella
pTDpelB-C_sfYFPTwinStrep Periplasmic space PelB signal sequence YFP Dammeyer
pmScarlet-I_peroxisome_C1 Peroxisome SRL mScarlet-I Gadella
pmScarlet-H_peroxisome_C1 Peroxisome SRL mScarlet-H Gadella
pmScarlet_peroxisome_C1 Peroxisome SRL mScarlet Gadella
mCherry-Peroxisomes-2 Peroxisomes Peroximal Targeting Signal 1 (PTS1) mCherry* Davidson
pcDNA3.1-KozATG-myc-dL5-2XG4S-mCer3-SKL Peroxisomes SKL tripeptide, peroxisome transport signal dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez

*Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein Collection.

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Cytoskeletal Elements

Browse or search the table below to find markers of cytoskeletal elements.

Plasmid Structure Targeting Gene/Signal Fluorescent Protein PI
pCAG-mGFP-Actin Actin Filaments beta-actin EGFP Yasuda
pLifeAct-mTurquoise2 Actin Filaments LifeAct mTurquoise2 Gadella
GFP-UtrCH Actin Filaments utrophin (aa# 1-261) GFP Bement
pIRESneo-EGFP-alpha Tubulin Microtubules alpha-tubulin EGFP Wadsworth
pmTurquoise2-Tubulin Microtubules alpha-tubulin mTurquoise2 Gadella
mCh-alpha-tubulin Microtubules alpha-tubulin mCherry Voeltz
pPAmCherry-b-actin Actin Filaments beta-actin PAmCherry1 Verkhusha
mCherry-Lifeact-7 Actin Filaments LifeAct mCherry* Davidson
pDEST/LIfeAct-mCherry-N1 Actin Filaments LifeAct mCherry Shaw
pEGFP Centrin2 Centrioles (dependent on cell cycle) Centrin-2 EGFP Nigg
pPAmCherry-a-tubulin Microtubules alpha-tubulin PAmCherry Verkhusha
EMTB-3XGFP Microtubules ensconsin GFP Bement
EMTB-mCherry Microtubules ensconsin mCherry Bement
pLifeAct_mScarlet_N1 Actin Filaments LifeAct mScarlet Gadella
pEGFP PICH Centromeres (dependent on cell cycle) PICH EGFP Nigg
pEGFP-mNFM Intermediate Filaments (neuronal) Nefm EGFP Brown
pLifeAct-miRFP703 Actin Filaments LifeAct miRFP703 Verkhusha
pLifeAct_mScarlet-i_N1 Actin Filaments LifeAct mScarlet-I Gadella
pLifeAct_mScarlet-H_N1 Actin Filaments LifeAct mScarlet-H Gadella
pEGFP Cep170 C-term Centrioles (dependent on cell cycle) Cep170 EGFP Nigg
mEmerald-PLK1-N-16 Centrosome Plk1 mEmerald* Davidson
pVimentin-miRFP703 Intermediate Filaments (vimentin) vimentin miRFP703 Verkhusha
pKeratin-miRFP703 Intermediate Filaments (epithelial) Keratin miRFP703 Verkhusha
mCherry-Tubulin-C-18 Microtubules alpha-tubulin mCherry* Davidson
pDEST/N1-hEB1-GFP Microtubules EB1 GFP Shaw
pmiRFP703-Tubulin Microtubules alpha-tubulin miRFP703 Verkhusha
pEB3-miRFP703 Microtubules EB3 miRFP703 Verkhusha
pmScarlet-i_alphaTubulin_C1 Microtubules alpha-tubulin mScarlet-I Gadella
pmScarlet-H_alphaTubulin_C1 Microtubules alpha-tubulin mScarlet-H Gadella
pmScarlet_alphaTubulin_C1 Microtubules alpha-tubulin mScarlet Gadella
pEGFP-rNFM Intermediate Filaments (neuronal) Nefm EGFP Brown

*Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein Collection.

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Cell Junctions

Browse or search the table below to find markers of cellular junctions.

Plasmid Structure Targeting Gene/Signal Fluorescent Protein PI
pDEST/hCx43-EGFP-N1 Gap Junctions Connexin 43 EGFP Shaw
pDEST-mCherry-GJA1-20k-N1 Gap Junctions Connexin 43 / GJA1-20k mCherry Shaw
mCherry-Cx43-7 Gap Junctions Connexin 43 / GJA1-20k mCherry* Davidson
E-cadherin-GFP Adherens junctions E-cadherin GFP Stow
Paxillin-pEGFP Focal adhesions paxillin GFP Horwitz
pPaxillin-PSmOrange Focal adhesions paxillin PSmOrange Verkhusha
mEmerald-Paxillin-22 Focal Adhesions paxillin mEmerald* Davidson
mApple-VE-Cadherin-N-10 Tight Junctions VE-Cadherin mApple Davidson
pZyxin-miRFP703 Focal adhesions Zyxin miRFP703 Verkhusha

*Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein Collection.

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Secretory Pathway

Browse or search the table below to find markers of secretory pathway components.

Plasmid Structure Targeting Gene/Signal Fluorescent Protein PI
pEF.myc.ER-E2-Crimson Endoplasmic Reticulum ER retention signal E2-Crimson Glick
pmTurquoise2-ER Endoplasmic Reticulum ER retention signal mTurquoise2 Gadella
mCh-Sec61 beta Endoplasmic Reticulum Sec61 mCherry Voeltz
BFP-Sec61 beta Endoplasmic Reticulum Sec61 BFP Voeltz
BFP-KDEL Endoplasmic Reticulum BiP ER signal sequence BFP Voeltz
Rtn4a-GFP Endoplasmic Reticulum Rtn4a AcGFP Voeltz
ERoxBFP Endoplasmic Reticulum ER retention signal oxBFP Snapp
ERmoxGFP Endoplasmic Reticulum ER retention signal moxGFP Snapp
GolgiCFP Golgi eNOS(1-33) CFP Newton
pmTurquoise2-Golgi Golgi B4GALT1(1-61) mTurquoise2 Gadella
GalToxBFP Golgi GalT signal anchor transmembrane domain oxBFP Snapp
Clathrin-LCa-EYFP Clathrin-coated vesicles Clathrin, light polypeptide EYFP Zhuang
mRFP-Clc Clathrin-coated vesicles Clathrin, light polypeptide mRFP Helenius
mEmerald-Clathrin-15 Clathrin-coated vesicles Clathrin mEmerald* Davidson
DsRed-Rab5 WT Early endosomes RAB5A DsRed2 Pagano
mRFP-Rab5 Early endosomes RAB5A mRFP Helenius
mCh-Rab5 Early endosomes Rab5 mCherry Voeltz
BFP-Rab5 Early endosomes Rab5 BFP Voeltz
GFP-Rab5B Early endosomes Rab5B AcGFP Voeltz
GFP-Rab4B Early endosomes Rab4B AcGFP Voeltz
GFP-rab7 WT Late endosomes RAB7 GFP Pagano
DsRed-rab7 WT Late endosomes RAB7 DsRed Pagano
mCh-Rab7A Late endosomes Rab7a mCherry Voeltz
GFP-Rab7A Late endosomes Rab7a AcGFP Voeltz
GFP-rab11 WT Recycling endosomes RAB11A GFP Pagano
DsRed-rab11 WT Recycling endosomes RAB11A DsRed Pagano
Lamp1-YFP Lysosomes Lamp-1 YFP Mothes
Lamp1-RFP Lysosomes Lamp-1 RFP Mothes
mCherry-Lysosomes-20 Lysosomes LAMP1 mCherry* Davidson
pCMB-AUTr Autophagosome LC3(ATG8) mCherry Harrison
pEGFP-LC3 (human) Autophagosome LC3 EGFP Finkel
pLAMP1-miRFP703 Lysosomes LAMP1 miRFP703 Verkhusha
pmiRFP703-Clathrin Clathrin-coated vesicles Clathrin miRFP703 Verkhusha
pmScarlet-i_Giantin_C1 Golgi apparatus Giantin mScarlet-I Gadella
pmScarlet-H_Giantin_C1 Golgi apparatus Giantin mScarlet-H Gadella
pmScarlet_Giantin_C1 Golgi apparatus Giantin mScarlet Gadella
pCytERM_mScarlet-i_N1 ER CytERM mScarlet-I Gadella
pCytERM_mScarlet-H_N1 ER CytERM mScarlet-H Gadella
pCytERM_mScarlet_N1 ER CytERM mScarlet Gadella
pTag-RFP-C-h-Rab11a-c-Myc Recycling endosomes Rab11a TagRFP Johnson
pTag-BFP-C-h-Rab11a-c-Myc Recycling endosomes Rab11a TagBFP Johnson
pTag-RFP-C-h-Rab7-c-Myc Late endosomes Rab7 TagRFP Johnson
pTag-BFP-C-h-Rab7-c-Myc Late endosomes Rab7 TagBFP Johnson
pTag-RFP-C-h-Rab5a-c-Myc Early endosomes Rab5a TagRFP Johnson
pTag-BFP-C-h-Rab5a-c-Myc Early endosomes Rab5a TagBFP Johnson
pTag-RFP-C-h-Rab4a-c-Myc Recycling endosomes Rab4a TagRFP Johnson
pTag-BFP-C-h-Rab4a-c-Myc Recycling endosomes Rab4a TagBFP Johnson
pcDNA3.1-kappa-myc-dL5-2XG4S-mCer3-KDEL Endoplasmic reticulum (ER) KDEL dL5 FAP (fluorogen activating protein); mCerulean3 fusion Bruchez

*Fusions to other FPs can be found by searching the complete Michael Davidson Fluorescent Protein Collection.

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