Invitrogen
Goat anti-Rat IgG (H+L) Secondary Antibody, Alexa Fluor 350
A-21093
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Our lab studies various cell types in the mouse retina and this antibody is weak with too much background fluorescent to use for our cells.
Immunofluorescence
Fixed whole mouse retina
1:200 primary antibody, 2 hours at room temperature in staining buffer (or overnight at 4 degrees celsius)
Goat serum
1:250 secondary antibody, 1 hours at room temperature in staining buffer (or overnight at 4 degrees Celsius)
None
Fluorescent microscope
We tried various combinations of primary and secondary dilutions and incubation times, but this antibody isn't great for detecting our cells. The background fluorescence is very high and the signal on the primary cells is weak compared with other colors (alexa 488 with same primary Ab).
N/A
Shows up well against red and green antibodies
Weak signal, high background
Not a great antibody for our uses; other colors worked better.