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|---|---|---|---|
| 100 ul | $325.00 | Add to Cart |
mCherry is derived from proteins originally isolated from Cnidarians (jelly fish, sea anemones and corals), and is used as a fluorescent tracer in transfection and transgenic experiments. The prototype for these fluorescent proteins is Green Fluorescent Protein (GFP), which is a ~27kDa protein isolated originally from the jellyfish Aequoria victoria. The mCherry protein is derived from DsRed, a red fluorescent protein related to GFP isolated from so-called disc corals of the genusDiscosoma. DsRed is similar in size and properties to GFP, but, obviously, produces a red rather than a green fluorochrome. Several further cycles of mutation, directed modification and evolutionary selection produced mCherry, which has an excitation maximum at 587 nm and and emission maximum at 610 nm (4). We expressed the mCherry protein sequence shown in reference 4 in bacteria, purified out the mCherry and raised this rabbit polyclonal antibody. This was affinity purified and was found to stain a band of the expected size in HEK293 cells transfected with the pFin-EF1-mCherry vector designed to express mCherry. As shown below, the antibody does not stain any protein band in untransfected HEK293 cells. |
Images
Blot of crude extract of HEK293 cells transfected with pFin-EF1-mCherry vector in lane labelled “+”. The “-” lane is a blot of an equal amount of protein extract from untransfected HEK293 cells. MCA-1C51 binds a major band running at ~28 kDa corresponding to intact full-length mCherry. The two other bands are clearly processed forms of mCherry as they are not present in non-transfected HEK293 cells
HEK293 cells transfected with mCherry and visualized in red. The cells were stained with MO22140 in the green channel, and visualized using a confocal microscope. Transfected cells are yellow, showing overlap of the mCherry and the MO22140 antibody. Untransfected HEK293 cells do not express Cherry and do not stain with the antibody, but their nuclei can be visualized using a DNA stain (blue). Blot and transfected cells courtesy of the Semple-Rowland lab at the University of Florida.