Cell-permeable, yellow fluorescent dye with an NHS ester (succinimidyl ester (SE)) reactive group. NHS esters react with primary amines on proteins and are commonly used for conjugating dyes to proteins, antibodies amine-modified oligonucleotides etc.
Janelia Fluor® 549 is 2 x brighter than TMR and Cy3 in vitro and live-cell experiments. Suitable for super resolution microscopy (SRM) including techniques such as STED and dSTORM (both live and fixed cells). May also be used for cellular imaging when combined with the HaloTag or SNAP-tag self labelling systems and confocal microscopy.
Yellow dye for coupling to primary amine groups. Suitable for super resolution microscopy (e.g. dSTORM, STED), confocal microscopy, live cell imaging and flow cytometry.
Fig 1. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 1. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin (HB6491) antibody from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti-mouse antibody. For more details see protocols #1 and #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and Neurofilament L (HB7266) antibodies from Hello Bio which were visualised using a Janelia Fluor® 549 SE conjugated goat anti-mouse antibody and a DyLight 488 conjugated goat anti- rabbit antibody. For more details see protocols #1 and #3 in the application notes below.
Fig 3. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 3. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin antibody (HB6491) from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti- mouse antibody. For more details see protocols #1 and #2 in the application notes
Figure 4. Janelia Fluor® 549 conjugated secondary antibodies show superior antifade performance compared to those conjugated with DyLight 550
Fig 1. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 1. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin (HB6491) antibody from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti-mouse antibody. For more details see protocols #1 and #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and Neurofilament L (HB7266) antibodies from Hello Bio which were visualised using a Janelia Fluor® 549 SE conjugated goat anti-mouse antibody and a DyLight 488 conjugated goat anti- rabbit antibody. For more details see protocols #1 and #3 in the application notes below.
Fig 3. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 3. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin antibody (HB6491) from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti- mouse antibody. For more details see protocols #1 and #2 in the application notes
Figure 4. Janelia Fluor® 549 conjugated secondary antibodies show superior antifade performance compared to those conjugated with DyLight 550
Add 10% by volume of 0.75M Tris-HCl pH7.4 (to a final concentration of 75mM) to stop the conjugation reaction. Incubate for 10-15 minutes at room temperature in the dark with gentle mixing.
Use a desalting column, perform buffer exchange (following manufacture instructions) of antibody into PBS 0.05% sodium azide. This step also removes any unbound dye
This compound is light and temperature sensitive; exposure to light may affect compound performance. We therefore recommend storing the material in a freezer and protecting from light.
Shipping conditions
Ship on ice
Important
This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use
Yellow dye for coupling to primary amine groups. Suitable for super resolution microscopy (e.g. dSTORM, STED), confocal microscopy, live cell imaging and flow cytometry.
![Janelia Fluor® 549, SE [1811539-32-8] Chemical Structure](https://cdn.hellobio.com/media/catalog/product//h/b/hb7336.png "Janelia Fluor® 549, SE [1811539-32-8]")
