WB was performed on seven GST-fusion proteins containing different fragments of AML-ETO (1-7) with the ab at 1:1,000. The antibody raised against AML-ETO recognizes the ETO part (lane 2) of the fusion protein. C: WB was performed on nuclear extracts from KAS-6/1 cells (human myeloma cell line) and SKNO-1 cells (human acute myeloblastic leukaemia) with the ab against AML-ETO at 1:1,000. The location of AML-ETO and a presumed splice variant (missing 106 C-terminal amino acids) are indicated.
Determination of the antibody titer To determine the titer of the antibody, an ELISA was performed using a serial dilution of antibody against AML-ETO, crude serum and flow through in antigen coated wells. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:5, 250.