It is also a major component of the Alexander test for pollen staining.

It is often combined with other yellow dyes and used to stain erythrocytes in the trichrome methods.

Orange G can be used as an electrophoretic color marker to monitor the process of agarose gel electrophoresis, running approximately at the size of a 50 Base pair (bp) DNA molecule, and polyacrylamide gel electrophoresis.

(However, the apparent "size" of all these dyes varies according to the concentration of agarose in the gel and the buffer system used, so one should look up the appropriate reference before using the dyes to determine how far a gel has run.)

Despite its three ionizable groups, it shows only two colors in aqueous solution, brilliant orange in neutral and acidic pH or red in pH greater than 9.