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1) What is the immunogen of the antibody? Generated different stable cell lines with mutations in the transferrin protein (aa95, 188, 426, 517). Will these mutations affect antibody binding?
2) Will the antibody/assay work equally well with transferrin bound to bicarbonate (naturally occuring) and oxycholate? See lower singal with transferrin bound to oxycholate.
3) Is quantification affected by the amount of iron loaded onto the transferrin molecule?
Asked on Feb 27 2015
1) The antibodies in this kit have been raised against native transferrin and unfortunately we do not know precise epitopes. A monoclonal capture antibody and a polyclonal detector antibody is provided. Therefore we do not know if the mutations generated will be recognized.
2 and 3) We have not tested the effects of bicarbonate, oxycholate or iron on assay activity. The standard protein in the kit is native purified human transferrin. The kit works with this protein and with the human serum/plasma samples and has very good linearity of dilution. Therefore we expect that this antibody pair is compatible with normal levels of bicarbonate/oxycholate/iron bound to transferrin. However we do not know if abnormally high/low levels of any of these will affect the reactivity of the sample.
Karen BakerAbcam Scientific Support
Answered on Feb 27 2015