Characterization in Western and IHC

Specificity in Western Blotting

The composite Western blot below in Figure 1 shows the characteristic doublet of our synapsin pan antibody as well as three of our phospho synapsin antibodies, and demonstrates two very import aspects of antibody specificity. First, there are no cross-reacting bands labeled by the antibody in the rat brain homogenate. Second, phospho-specificity is definitively demonstrated by comparing the signal in untreated rat brain homogenate (left) to the same homogenate, but that has been treated with lambda and alkaline phosphatase prior to being run on the gel (right). Treatment with phosphatase had no effect on the pan antibody’s signal, but completely eliminated the immunolabeling of the phospho antibodies.

Synapsin Phosphospecificity in Western Blot
Figure 1

Immunohistochemistry

Now we turn to immunohistochemistry. Many of our antibodies have been tested in IHC. Figure 2 below shows staining of cultured neurons with anti-synapsin pan antibody in green (left), and of C57 mouse striatal cells with anti-synapsin Ser549 (right).

Synapsin antibody validated in IHC
Figure 2

Figure 3 below illustrates our pan-TH antibody’s extensive labeling in this photomicrograph of the retina (left). In contrast, a phospho-TH antibody selectively labels only the two amacrine cells in this light-stimulated retina example (right).

Tyrosine Hydroxylase antibody validated in IHC
Figure 3