Tools for Multi-Color Protein Imaging

The ability to image cells is of great use to researchers studying any aspect of cell biology. Cellular imaging allows researchers to further probe complicated cell pathways of both diseased and normal tissues.

As our understanding of cellular processes and dynamics has evolved into the post genomic era it is now widely accepted that most proteins do not work in isolation, and instead they work in unison as complexes of multiple proteins aggregated together.

For this reason, researchers must have the tools to study proteins as part of complexes, rather than in their individual state.

If not equipped properly research involving multi-color imaging is complicated. It can be difficult to find primary antibodies which are labelled appropriately and phenomena such as cross species reactivity in secondary antibodies can give erroneous results.

To help researchers get past these challenges and the most out of multi-color imaging Abcam supply a range of research tools.

Multiplex Imaging with Alexa Fluor® Secondary Antibodies

The Alexa Fluor® range of conjugated secondary antibodies provide a low background and high staining with a high reliability, as proven by internal testing.

Abcam can also provide a wide range of antibodies which have been pre-adsorbed to minimize cross species reactivity. This makes the Alexa Fluor® range perfect for multi-color imaging.

  • Red staining - primary antibody mouse anti-disabled 2.  Secondary antibody goat anti-mouse IgG - H&L (Alexa Fluor® 594) pre-adsorbed (ab150120).
  • Green staining - primary antibody rabbit anti-laminin.  Secondary antibody goat anti-rabbit IgG - H&L (Alexa Fluor® 488) pre-adsorbed (ab150081).
  • Blue staining - nuclei counter stained with DAPI.

Tools for staining cellular organelles

Abcam provide a range of dyes for the staining of organelles and other subcellular structures. The staining follows a simple method – all researchers must do is prepare the cells and apply the dye. Following incubation, the cells are then ready for analysis.

Cellular organelle Product
Actin filaments CytoPainter F-actin available in blue, green, orange, and red.
Endoplasmic reticulum CytoPainter ER - available in green and red.
Golgi CytoPainter Golgi
Lysosomes CytoPainter lysosomal
Nucleus CyTRAK Orange™
DRAQ5™
DRAQ7™
Other nuclear stains
Mitochondria CytoPainter mitochondrial staining kit

Image: courtesy of Dr Shaohua Li, UMDNJ-Robert Wood Johnson Medical School.

Image: courtesy of Dr Shaohua Li, UMDNJ-Robert Wood Johnson Medical School.

  • Red - nuclei counter stained with DRAQ7™ (ab109202).
  • Green - primary antibody rabbit anti-laminin alpha.  Secondary antibody goat anti-rabbit IgG H&L FITC (ab97050).
  • Blue - F actin was stained with CytoPainter (ab112124).

Direct Labelling with Antibody Conjugation Kits

When undertaking research involving multi-color imaging, sometimes researchers will have to use two antibodies which have been raised in the same host.

In this situation if a secondary antibody is introduced then it will be impossible to differentiate between the two different primary antibodies. To avoid this problem Abcam can provide labelling kits for the direct labelling of primary antibodies with a fluorescent probe. The kits require only 30 seconds of work to apply and dues can be provided in wavelengths from ultraviolet to far IR.

Preserving Sample Fluorescence

When using an immunofluorescent imaging method, the loss of fluorescence whilst a sample is stored can present difficulties. However, using Abcam’s fluoroshield mounting media fluorescence can be preserved. The fluoroshield has the combined effect of slowing the photobleaching of dyes and retaining sample fluorescence.

Image: courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical school.

Image: courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical school.

About Abcam

Abcam is a global life sciences company providing highly validated antibodies and other binders and assays to the research and clinical communities to help advance the understanding of biology and causes of disease.

Abcam’s mission is to serve life scientists to help them achieve their mission faster by listening to their needs, continuously innovating and improving and by giving them the tools, data and experience they want. Abcam’s ambition is to become the most influential life science company for researchers worldwide.


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Last updated: May 2, 2019 at 9:39 AM

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