Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil: reactive)
Specification
Tonsil: reactive
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: 21°C
Other product details
Dilution
1/1000
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 21°C · Diluent: TBS/BSA/azide
Secondary antibody
Name
Non-Abcam antibody was used: Anti rabbit IgG
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Host species: Goat
Clonality: Polyclonal
Conjugation: Biotin
Dilution
1/200
Additional data
Additional Notes
I rate this as "Good" because I appear to obtain a similar positivity to that of ab76496 ( image of human tonsil in datasheet).
The dilution factor for human tissue appears to be much lower than that for rat and mouse, in my hands.
The submitted image shows clear cytoplasmic positivity in a subset of germinal centre cells: the cells in the mantle zone also appear weakly positive but it is difficult to see this, in submitted image.
A lower dilution factor of primary Ab. would show these cells more clearly but,at the dilution factor I used, there was intense positivity of the serum in the blood vessels ( indicated by red arrowheads - I do not know if this is specific/non-specific).
NB: I blocked endogenous peroxidases using 2% H2O2, for 15 minutes.
The near sequential section that I immunostained for pFAK, at the
same time, did not give this serum positivity.
The dilution factor for human tissue appears to be much lower than that for rat and mouse, in my hands.
The submitted image shows clear cytoplasmic positivity in a subset of germinal centre cells: the cells in the mantle zone also appear weakly positive but it is difficult to see this, in submitted image.
A lower dilution factor of primary Ab. would show these cells more clearly but,at the dilution factor I used, there was intense positivity of the serum in the blood vessels ( indicated by red arrowheads - I do not know if this is specific/non-specific).
NB: I blocked endogenous peroxidases using 2% H2O2, for 15 minutes.
The near sequential section that I immunostained for pFAK, at the
same time, did not give this serum positivity.
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.
MR. Carl Hobbs
Verified customer
提交于 Jul 19 2010