Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Skin)
Permeabilization
Yes - 0,3 % Triton X
Specification
Skin
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Perfusion fixed with paraformaldehyde and picric acid
Other product details
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: TBS
Dilution
1/50
Secondary antibody
Dilution
1/200
Name
Non-Abcam antibody was used: Dianova Donkey anti Rabbit Cy3
Host species: Donkey
Clonality: Polyclonal
Conjugation: Cy3
Host species: Donkey
Clonality: Polyclonal
Conjugation: Cy3
Additional data
Additional Notes
Involucrin (Cy3/Red), Avidin (FITC-Avidin/Green), Nucleus (DAPI/Blue)
Antibody shows unclear and very weak staining in epidermis, not comparable to expression and intensity shown in literature (Yamamoto et al. 2008)
Abcam response
Thank you for your reveiw of this previously untested application, IHC of PFA/picric acid-fixed frozen tissue. The IHC data we have received from customers for formalin-fixed, paraffin-embedded tissue suggests that a heat-mediated antigen retrieval may be necessary before staining to develop a strong signal.
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 Aug 12 2014