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Figure 1. Determination of the titer To determine the titer, an ELISA was performed using a serial dilution of the Diagenode antibody directed against human MBD4 (Cat. No. CS-087-100) in antigen coated wells. 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 1:15,000.
Figure 2. Western blot analysis using the Diagenode antibody directed against MBD4 Figure 2A: Human osteosarcoma cells (U2OS) were transfected with an expression vector for TY1-tagged MBD4. The presence of TY1-MBD4 in the cell lysates was demonstrated by western blot analysis with the Diagenode antibody against the TY1-tag (Cat. No. MAb-054-050) (lane 1) and with the antibody against MBD4 (Cat. No. CS- 087-100) (lane 2), diluted 1:2,000 in TBST containing 3% milk powder. Figure 2B: Western blot was performed on nuclear extracts from the U937 human leukemic monocyte lymphoma cell line (40 μg) with the antibody against MBD4, diluted 1:2,000 in TBST containing 3% milk powder. A molecular weight marker (in kDa) is shown on the left. The location of the protein of interest is indicated on the right.
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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