Immunohistochemistry (Frozen sections) abreview for Anti-Otx1 + Otx2 antibody - ChIP Grade

Average
Abreviews
Application
Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Mouse Tissue sections (E8.5 mouse embryo brain section)
Specification
E8.5 mouse embryo brain section
Permeabilization
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Fixative
Paraformaldehyde

Other product details

Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 1% BSA + 5% donkey serum + PBST
Dilution
1/300

Secondary antibody

Name
Non-Abcam antibody was used: donkey anti-rabbit 555 (Invitrogen)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor­ 555

Additional data

Additional Notes
I see specific nuclear staining in the brain of E8.5 embryos using this antibody. But I have given the antibody 3 stars, because there is a lot of unspecific membrane staining in the posterior of E8.5 sections, which I have difficulty to get rid of. I have tried to use glycine or a commercial primary dilution buffer to reduce the unspecific staining, but neither worked.

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Submitted Nov 05 2013

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