Hrp conjugated goat anti mouse igg light chain specific antibody

Immunoprecipitation was performed^{67 (link)} using the synaptosomal fraction from the pooled dissected tissues from the dentate gyrus of 4–5 mice (for Figures 1b and Supplementary Figure 4) or pooled whole cell lysate from the hippocampus of 2–3 mice (for Figure 8a–c) were used for the immunoprecipitation experiments. The lysates (1–2 mg of protein) were precleared with Protein G Plus Agarose beads (Thermo Scientific) and with 2–5 μg of anti-α-tubulin (Sigma), anti-KIF5 (Millipore), or normal mouse IgG antibodies (Santa Cruz Biotechnology), followed by incubation with Protein G Plus Agarose beads. The beads were washed 5 times with RIPA buffer. Proteins were eluted with a sample buffer containing 1% SDS. Western blot analyses of the immunoprecipitated anti-α-tubulin, anti-KIF5, and normal mouse IgG samples and the input (from the precleared step) samples were performed with anti-stathmin, anti-GluA2, anti-KIF5, anti-α-tubulin, or anti-actin antibodies as described above. HRP-conjugated horse anti-mouse IgG antibody (Cell Signaling), HRP-conjugated goat anti-rabbit IgG antibody (Cell Signaling), or HRP-conjugated goat anti-mouse IgG light chain specific antibody (Jackson ImmunoResearch) were used as secondary antibody.