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Inquiry: Dear Abcam, I have a novel protein that has been fused to Fc. I was wondering if your easylink antibody kits would be able to create a protein:fluorochrome conjugate if there is an N-terminal Fc fusion?
Asked on Aug 22 2013
Thank you for your enquiry.
The EasyLink conjugation kits are designed so that they can be used to label proteins other than antibodies. In this case you should be able to label the Fc fused protein in the same way as labeling an antibody. However as the size of their protein is likely different to that of an average antibody, please find advice we give in these situations below:
As the protocols provided were optimised for labeling IgGs, we would recommend to adjust the amount of material added to the label vial to allow for molecular weight difference. This should be done without changing the volume added to the vial, as this could affect the conjugation efficiency. As a rough guideline, we would recommend changing the amount of material proportionally to the size difference with IgGs. An average IgG is about 160kD, therefore for a target that is ½ the size of an antibody (about 80kD), add ½ as much to the vial.
Finally, please note the usual EasyLink requirements will still apply: the target should be purified, in a suitable amine free buffer and not contain any additives such as Azide, BSA, Tris or Glycine.
I hope that this helps. If you have any additional questions please do not hesitate to contact me.
Sam WasherAbcam Scientific Support
Answered on Aug 22 2013