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Figure 1. Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human MBD3 (cat# CS-079-100). The wells were coated with the peptides used for immunisation of the rabbit. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:13,890.
Figure 2. Western blot analysis using the Diagenode antibody directed against MBD3 Figure 2A: Western blot was performed on nuclear extracts from a Human leukemic monocyte lymphoma cell line (U937 NE, 60 μg) with the Diagenode antibody against human MBD3 (Cat. No. CS-079-100), diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest is indicated on the right. Figure 2B: TY1-tagged MBD3 (TTE-MBD3) was over-expressed in osteosarcoma cells (U2OS). Nuclear extracts were prepared and separated by electrophoresis. An antibody directed against the TY1 tag or the antibody against MBD3 (diluted 1:2,000 in TBS-Tween containing 5% skimmed milk) were used to detect TTE-MBD3 (lane 1 and 2, respectively). The location of tagged MBD3 is indicated with an arrow.
WB Western blot : The quality of antibodies used in this technique is crucial for correct and specific protein identification. Diagenode offers huge selection of highly sensitive and specific western blot-validated antibodies.
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