Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Brain)
Specification
Brain
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH, Neutral, Low pH
Permeabilization
Yes - 0.25% Triton
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Other product details
Dilution
1/100
Incubation time
15 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 2.5% Horse Serum
Secondary antibody
Name
Non-Abcam antibody was used: Vector
Host species: Horse
Clonality: Polyclonal
Conjugation: HRP polymer
Host species: Horse
Clonality: Polyclonal
Conjugation: HRP polymer
Additional data
Additional Notes
Stain worked very well for frozen sections, but we could not get it to work on paraffin sections. Tried multiple antigen retrieval methods (pH 6.0/pH 7.4/pH 10.0 HIER, proteinase K, trypsin) as well as no antigen retrieval. We also tested different permeabilization methods (triton X-100 at 0.25% or 0.5% or 1%, SDS, tween-20 in all wash buffers and incubation buffers) to no avail. Different concentrations of antibody were tested also.
At first, we thought that it might have been a tissue preparation issue, but a different antibody from another vendor worked beautifully with high pH antigen retrieval. Perhaps this antibody just doesn't work well with our preparation methods. Staining was always non-positive, giving us clean tissue despite prolonged development with DAB.
At first, we thought that it might have been a tissue preparation issue, but a different antibody from another vendor worked beautifully with high pH antigen retrieval. Perhaps this antibody just doesn't work well with our preparation methods. Staining was always non-positive, giving us clean tissue despite prolonged development with DAB.
Abcam response
Our Scientific Support team is currently investigating this negative Abreview, and we will resolve the issue in accordance with our Abpromise.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
Abcam user community
Verified customer
提交于 Nov 05 2012