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We have been trying to work out the nephrin antibodies (ab85379, and
ab58968) from Abcam for the past couple weeks. We have attempted
paraffin embedded sections, frozen sections, with both 4% PFA and
5% Formalin fixation methods (for both paraffin and frozen sections).
We have also tried multiple citrate buffer antigen retrieval solutions.
In the end, we couldn't get a specific staining in the glomerular
basement membrane. The most successful attempt using a signal enhancer
kit produced a positive stain in the tubules (this is not where
Nephrin is expressed).
We are considering abandoning this antigen and trying to stain for
another glomerular marker.
As loyal Abcam users, we thought to attempt to try an Anti-NPHS2
antibody (ab50339) using our IHC protocol on paraffin embedded tissues.
We would attempt cadaveric rat, pig, and human kidney sections.
Would we be able to get this antibody to work out?
Asked on May 03 2012
Thank you for your email.
I am sorry to hear that these antibodies are not working as expected.
As for ab58968, we had replaced this antibody free of charge with ab85379. Since ab85379 was not tested in rat, sheep, and pig tissue, I had sent you a testing discount code. The code (xxx) is still valid until its expiration date of xxx.
Which species did you try ab85379 on?
As for ab50339, it has only been tested in WB, IHC-Fr, ICC/IF, but not paraffin embedded sections. It might work on paraffin sections, but we have no data to confirm this as it was not yet tested.
I'd be happy, though, to give you a testing discount code for trying it in IHC-P. Would you use it on the same species as ab85379?
I look forward to hear back and assist you further.
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Answered on May 03 2012