Explore research tools to identify protein subcellular localization.
High-resolution imaging allows you to track the location and movement of proteins within the cellular environment. To ensure proper interpretation of these experiments, it is necessary to confirm whether the protein is actually located in the subcellular environment we expect.
Two approaches can be used to confirm the subcellular localization of a protein: organelle-specific antibodies and organelle stains.
Organelle stains can be used as counterstains to help identify the location of specific proteins and targets of interest within the cell, while antibodies against proteins associated with a specific organelle can lead to a better understanding of cellular function.
For nuclear staining, you have several options, including the highly success DRAQ staining.
Easy to implement in your current staining protocols
Suitable for proliferating and non-proliferating cells
Maybe used in most cellular or tissue samples
Minimal photobleaching allows longer exposure times
When might a Cytopainter dye work best for you?
Working with live cells. The currently available range of antibodies can only be used on fixed cells, not when studying a time-lapsed event in live cells. Most organelle dyes will stain subcellular compartments in live cells as well as being retained within said compartments after a fixation step.
This versatile characteristic means that they can also be used for co-staining experiments that use antibodies.
Studying multiple proteins. The number of proteins that can be studied using antibodies is limited by the number of species in which fluorescence-linked antibodies are available.
Organelle dyes bypass this limitation as they are chemical compounds, making them an excellent alternative to antibodies when the experiment requires multiple targets.
Morphology/distribution studies. When studying disease models and mutated cells, it is possible that the marker protein will not be localized in the subcellular compartment where it should be, or that the morphology of the organelle might be compromised.
Organelle stains will stain the subcellular structures as long as they are intact, even if its morphology has been changed.