I am using one antibody from cell signaling, which seems to stain projections from the granular zone and stains in between the BRDU positive cells, and i'm using a different doublecortin antibody from Santa Cruz which stains inside the cells and appears to colocalize with the BRDU. In the images I attatched, the Doublecortin is green and the BRDU is red. These images are taken from P6 mice. The first three images correspond to the antibody from Cell signaling, and the last three are from Santa Cruz.
Hi Tatyana,
DCX is a precursor neuronal marker, so you need to see neurons with their progenitors.
I am using IgG anti DCX from Abcam, that works fine for me.
none of them looks really specific. You should see young neurons in a sparse labelling only.
What works is st cruz sc-8066 1:200 dilution (I use vibratome sections), or millipore AB2253 1:500
Your cell signaling antibody looks more specific but the background labeling is high which makes it hard to visualize the projections. The santa cruz antibody is not good here. Good luck.
Thank you so much for all your input! I will try increasing my antibody concentration from 1:750 to 1:200, and maybe get another company antibody. Would it make a difference that my mice are P6? Most of the staining is done in adult mice, which might indeed have a smaller population of young neurons. P6 is the peak of neuroproliferation in mice.
Tatyana I agree with Kate, It seems more specific the cell signaling antibody, because the signal is in the citoplasm and it is in the inner part of the GCL, where newborn adult granule cells are located and have DCX until 21 days post born.
Good luck with your immuno.
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