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Asked on Jul 17 2012
Sorry for the delay in getting back to you.
I now have some further information to share with you in regards to the antibody ab89792. This antibody is a mouse monoclonal which has been raised against a recombinant fragment of the human LIFR protein, from residues 45 to 833 (SwissProt reference P42702). This recombinant protein was also used as the positive control when verifying this antibody, giving a band at approximately 110 kDa.
I think the bands observed by your colleague are as would be expected. The full length protein is predicted to be 124 kDa, but this does not take into account the glycosylation which the protein undergoes. I have found reference to the protein being detected at ˜160 kDa in murine samples where a size of ˜123 kDa is predicted from the SwissProt reference P42703:
Unfortunately, I have had some difficulty intrying to find further publications where the size of the receptor detected through Western blotting is mentioned. However I think the band at 150 kDa could well be the glycosylated, membrane bound LIFR protein.
The band observed at 100 kDa could correspond to the soluble LIFR protein. The soluble protein encompasses residues 1-833. I was again unable to find direct references to the size of protein which has been detected but the size of the protein sequence alone would give 94 kDa. As I think the N-terminal signalling sequence (residues 1-45) is sometimes cleaved from the soluble protein this would reduce this size somewhat, but again with glycosylation you can expect the size to increase. I therefore think a size around 100-110 kDa would be expected.
I am sorry but I will not be able to provide you with a blocking peptide as the antibody is raised against the recombinant protein and the exact binding epitope of the antibody is not known. If you would be interested, I could offer you a testing discount on trying out other anti-LIFR antibody (ab101228). This antibody has not been tested with human samples as yet but as the immunogen used shares 100% homology with the human protein I would expect the antibody to be able to detect the human protein. Withthis antibody I could enquire if we would be able to sell the blocking peptide as well as the antibody if you would like.
I think we have discussed this offer before but here is the link for your information again:
I hope this information has been of help, if you have any further questions, please do not hesitate to contact us again.
Answered on Jul 17 2012