Figure 4 : Line-blot for qualitative analysis of anti-BDNF antibodies specificity.

From: A method for reproducible measurements of serum BDNF: comparison of the performance of six commercial assays

Figure 4

(A) The antibodies from each ELISA kit were tested for specificity against pro-BDNF or mature BDNF. The BDNF standards blotted were commercial pro-BDNF (Alomone; 10 pg/lane), mature BDNF (1 and 2 from Alomone and Sigma, respectively; both 1000 pg/lane) and the standard BDNF protein included in each kit (Aviscera-Bioscience and Biosensis: 10 pg/lane; Millipore-ChemiKineTM, Millipore-Milliplex®- and R&D System-Quantikine®: 100 pg/lane; Promega-Emax®: 1000 pg/lane). BSA (1000 pg/lane) was used as a negative control. The mouse monoclonal anti-BDNF antibody, (1:1000; Sigma) was tested as a control. (B) Central region of the same blot shown in A, from an overexposed film to better visualize the reactivity against pro-BDNF. (C) Reactivity of antibodies from Biosensis, Promega-Emax® pAb and Sigma on a dot blot in which the same quantity of pro-BNDF and mature BDNF were spotted (100 pg each). Each antibody from the ELISA kits was used at the dilution suggested by the manufacturer’s instructions. mAb: Promega-Emax® monoclonal capture antibody for plate coating. pAb: Promega-Emax® polyclonal detection antibody.