The Golgi complex is the organelle where macromolecules, including proteins, lipids, and carbohydrates, are prepared for either secretion from the cell or use by other organelles within the cell. Many of the cell-permeant probes that selectively stain the Golgi complex have applications in lipid metabolism and trafficking studies and can also be used for measuring rates of lipid synthesis. Both antibodies and lectins can be used to target the Golgi complex in fixed cells, and fluorescent proteins allow for tracking in live cells. Three different Molecular Probes® technologies are useful for imaging the Golgi complex in live or fixed cells.

CellLight® reagents for Golgi complex

Use CellLight® reagents when you want to label the Golgi complex in live cells to follow the dynamics of intracellular behavior. CellLight® proteins can be multiplexed with other fluorescent probes for analysis in live cells or after fixation for colocalization studies. These ready-to-use constructs employ a human Golgi-resident enzyme, N-acetylgalactosaminyltransferase-2 (12.6 kDa), to target the Golgi complex with fluorescent fusion proteins. Introducing CellLight® reagents involves a simple transfection step, and they work like cell stains with minimal toxicity or chemical disruption.

CellLight® reagentsEnlarge image

Ceramide dyes for live cells

Fluorescent ceramides are used extensively to study lipid metabolism and trafficking. They provide selective staining for the Golgi complex in live cells and can also be fixed. NBD C6-ceramide is weakly fluorescent in aqueous solvents, and its fluorescence increases in a nonpolar environment. The BODIPY® dye conjugates are brighter and more photostable, with less environmental sensitivity. Complexing fluorescent lipids with bovine serum albumin (BSA) eliminates the need for organic solvents to dissolve the lipophilic probe—the BSA-complexed probe can be directly dissolved in water and loaded into live cells.

BODIPY® FL C5-ceramideEnlarge image

Conjugates for fixed-cell staining

The Golgi complex can be labeled in fixed cells using fluorescent conjugates of lectins such as HPA, or antibodies directed against Golgi-associated proteins such as golgin-97. HPA lectins selectively bind to terminal α-N-acetylgalactosaminyl residues—intermediate sugars added in O-linkage to serine and threonine residues in cis-Golgi cisternae. The anti–golgin-97 monoclonal antibody recognizes the Golgi complex of most vertebrate species, specifically at the peripheral membrane protein. We offer anti–golgin-97 as an unlabeled primary antibody, and HPA lectin as a conjugate of Alexa Fluor® 647 or Alexa Fluor® 488 dye.

Golgin-97 Monoclonal AntibodyEnlarge image
Selection guides
 
Readout
 Expression of fluorescent fusion protein
Target
 Protein targets Golgi complex
Common filter set
FITC
TRITC
Labels
GFP
RFP
Ex/Em (nm)
488/520
555/584
Signal-to-noise ratio
Photostability
Multiplexing
Yes
Yes
Live cells
Yes
Yes
Fixed cells
No
No
Fixable
Yes
Yes
Platform
Imaging 
Imaging 
Format
1 mL
1 mL
Cat. No.
 
Readout
Live-cell Golgi complex staining  
Target
 Targets lipids in Golgi complex
Common filter set
FITC
Texas Red®
FITC
Labels
BODIPY® FL
BODIPY® TR
NBD
Ex/Em (nm)
504/511
587/615
466/536
Signal-to-noise ratio
Photostability
Multiplexing
Yes
Yes
Yes
Live cells
Yes
Yes
Yes
Fixed cells
No
No
No
Fixable
Yes
Yes
Yes
Platform
 Imaging
 Imaging
 Imaging
Format
5 mg
5 mg
1 mg
Cat. No.
 
Readout
 Fixed-cell Golgi complex staining
Target
 Golgi membrane protein
α-N-acetylgalactosamine residues  
Common filter set
NA
FITC
Cy®5
Labels
Requires secondary detection
Alexa Fluor® 488
Alexa Fluor® 647
Ex/Em (nm)
NA
496/519
650/665
Signal-to-noise ratio
NA
Photostability
NA
Multiplexing
Yes
Yes
Yes
Live cells
No
No
No
Fixed cells
Yes
Yes
Yes
Fixable
No
No
No
Platform
 Imaging
 Imaging
 Imaging
Format
100 μg
1 mg
1 mg
Cat. No.
For Research Use Only. Not for use in diagnostic procedures.