Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-B Raf antibody [EP152Y]

Poor
Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (saggital sections of cerebellum and cortex)
Specification
saggital sections of cerebellum and cortex
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C

Other product details

Dilution
1/200
Incubation time
16 hour(s) and 0 minute(s) · Diluent: TBS/BSA/azide

Secondary antibody

Name
Non-Abcam antibody was used: anti rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Dilution
1/300

Additional data

Additional Notes
Submitted composite image shows (top) rat cerebellum: some Purkinje cells have strong cytoplasmic staining of the cell bodies but most are weakly staining or negative. Also, some primary processes are weakly positive. At higher concentrations of primary Ab, the molecular layer is weakly positive.
In the lower image, the deep cortex and corpus callosum is seen: there is an indistinct "neuropil" positivity.
Increasing the concentration reveals no additional structures.
I was looking at the svz of the lateral ventricle (site of olfactory neurone replacement generation) but was unable to detect any positivity in this region, hence submitting images of what I consider to be the "most positive" regions.
Abcam response
Many thanks for your feedback. This antibody has been previously tested in IHC-P using human tissue. Your negative result may reflect poor accessibility of the epitope recognized by the antibody in rat tissue. It may be useful to try different fixatives and/or antigen retrieval method. We welcome feedback from other customers who have tried this antibody for IHC-P in rat tissues.

Mr. Carl Hobbs

Verified customer

Submitted Feb 03 2009

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