Fig 1: Detection and quantification of BDNF in mouse hippocampus with different monoclonal antibodies. A, BDNF-IR using mAb#9 antibody in P21 WT C57Bl6/J hippocampus. CTIP-2 expression highlights granule cells in the dentate gyrus and pyramidal projection neurons in cornu ammonis (CA) 1-3. B, BDNF-IR in hippocampal mossy fiber projections of 8 week male WT C57Bl6/J versus NFL-Cre BDNFfl/ko hippocampus, using mAb#9. C, Specificity of 4 independent BDNF antibodies in CA3 mossy fiber terminals of WT C57Bl6/J (left column) and NFL-Cre BDNFfl/ko (right column). Clone 3C11 failed to detect endogenous BDNF. 3B2 produced high background in BDNF depleted hippocampal sections. The signal-to-noise ratio appeared best with mAb#9 and also with 4C8, which showed intense BDNF-IR. D, Double staining of BDNF and Myc in hippocampus of 8 week male BDNF-myc mice. Myc was costained with two independent polyclonal myc antibodies (Abcam, AB9106; Santa Cruz Biotechnology, SC789). E, BDNF protein levels in different CNS mouse brain areas, as determined by Western blot (top) with the 3C11 antibody and ELISA (bottom) using mAb#9 and mAB#1. P21 male WT C57Bl6/J (white bars) mice were compared with male NFL-Cre BDNFfl/ko mice (black bars) with bdnf gene recombination in most pyramidal neurons. Data are mean ± SEM. Hip, Hippocampus; CTX, anterior cortex; STR, striatum; Cereb, cerebellum (n = 3 for WT Hip, n = 5 for CTX, Cereb, STR, n = 2 for NFL Cre BDNFfl/ko Hip, CTX, STR, Cereb). Raw data are provided in Extended Data Figure 1-1 and Table 2. Image type: A, 2D merged single-plane image; B, D, maximum intensity projection; C, average intensity projection. Scale bars: B, D, 200 µm; C, 100 µm.
Fig 2: Commercial Pgap2 antibody does not detect overexpressed or endogenous PGAP2.10μg lysate from overexpression of mouse Pgap2-myc tagged clone (Origene #MR203189) in WT (lane 2) or Pgap2 KO 293T (lane 3) or lysate from mock transfection of WT (lane 4) or Pgap2 KO 293T (lane 5) probed with 1:500 Rb αPGAP (Thermo #PA5-64091, Green). (A) 10μg lysate from overexpression of mouse Pgap2-myc tagged clone in WT (lane 2) or Pgap2 KO 293T (lane 3) or lysate from mock transfection of WT (lane 4) or Pgap2 KO 293T (lane 5) probed with 1:1000 Rb αMyc (Abcam #ab9106, Green) or Ms αTub (Red) (B).
Supplier Page from Abcam for Anti-Myc tag antibody