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I know that your company send me the details how to refund on off the antibodys. But I can’t do it via this way because it doesn’t fit with our computers here.
Answer ur question:
1) WB, high background, now Bands in the Western Blot
IF, high background and none specific Binding
FACS, only background but now speciffic binding
2) Antibody dilutions
ab93909 2µg/ml to 10µg/ml FACS, IF and WB
ab128487 1: 50 bis 1:125 FACS, IF and WB
3) 20µg of Protein been loaded tot he WB like all off my WB looks like
4) I tried my old protocoll which I send u last time, we fixed the cells and +/-permeabilisation I tried too
5) The second antiboys are perfect because i use them for other IF, WB and FACS too and there they are looking perfect
Asked on Nov 07 2012
Thank you for your reply and the additional information.
Regarding the refund, you would need to let your purchasing department know the information I had sent and they then should contact us so that the refund can be issued to you. I'm certain there is a way this can be arranged after your institution gets in touch with us.
Regarding the other 2 antibodies:
Your protocol in general looked fine and the antibody dilution used are more concentrated than the suggested range.
You mentioned that you used Hela cells and I am wondering - since 3 anti-thrombopoietin antibodies seem not to be working - if this protein is expressed in Hela cells.
It is mainly present in megakaryocytes and megakaryocytes-derived cells.
The protein was shown to be present also in MCF7 cells, liver tissue, HepG2 cells.
What other cell types have you tried, besides Hela cells? Would you be able to try any of the positive controls?
I look forward to hear back from you and help you further.
Answered on Nov 07 2012