Anti-mCherry Antibody (A85304)

Immunohistochemistry analysis of formalin fixed paraffin embedded tissue from a mouse transgenically expressing a Cre dependent oxytocin-Tdtomato construct in brain tissue. TdTomato is very closely related to mCherry, so Anti-mCherry Antibody (A85304) recognizes both proteins. The Anti-mCherry Antibody (A85304) at a dilution 1:10,000 detected in DAB (brown) following the ABC method and citra buffer heat retrieval. Counterstained with Hematoxylin (blue). The Anti-mCherry Antibody (A85304) specifically detects as expected the soma and axons of Cre expressing neurons.

HEK293 cells transfected in the same way and viewed in the confocal microscope. Most HEK293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain. Cells which are transfected with Cherry are bright red. Staining with Anti-mCherry Antibody is shown in Green. Green antibody staining is only seen in cells which express Cherry, as expected, and the superimposition of the green and red results in an orange signal. Interestingly stronger Cherry staining is seen in the nucleus, possibly due to degradation of some Cherry molecules to release the low molecular weight Cherry fluorochrome.

Immunofluorescent analysis of HEK293 cells transfected with mCherry and stained with Anti-mCherry Antibody (A85304), viewed in a confocal microscope. Most HEK293 cells are not transfected so only the nucleus of these cells can be visualized with a blue DNA stain. Cells which are transfected with mCherry are bright red, and staining with Anti-mCherry Antibody (A85304) is shown in Green. The green antibody staining is only seen cells which express mCherry, as expected, and the superimposition of the green and red results in an orange signal. Interestingly stronger mCherry staining is seen in the nucleus, possibly due to degradation of some mCherry molecules releasing the low molecular weight mCherry fluorochrome.

Western blot analysis of HEK293 cell lysates using Anti-mCherry Antibody (A85304) at a dilution of 1:2,000, (green), and rabbit pAb to GAPDH, RPCA-GAPDH at a dilution of 1:5,000 (red). The lanes contain: [Lane 1] protein molecular weight standard, [Lane 2] untransfected HEK293 control cells, [Lane 3] HEK293 cells transfected with pCI-Neo-mod vector expressing two tdTomato protein domains, [Lane 4] HEK293 cells transfected with pCI-Neo-mod vector expressing one mCherry-HA protein domain, and [Lane 5] HEK293 cells transfected with pCI-Neo-mod vector expressing one GFP domain. The Anti-mCherry Antibody (A85304) antibody recognizes tdTomato and mCherry proteins revealing major bands at about 60kDa and 30kDa, but does not recognize GFP. The red band at 37kDa corresponds to GAPDH protein here used as a loading control.