Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain)
Antigen retrieval step
None
Permeabilization
No
Specification
Brain
Blocking step
10% NGS-2% Horse Serum in 0.1% Triton X-100-PBS as blocking agent for 1 hour(s) and 0 minute(s) · Temperature: 24°C
Fixative
Paraformaldehyde
Other product details
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.1%TritonX-100-PBS
Dilution
1/100
Additional data
Additional Notes
Rats were perfused with cold PBS, cold 4% PFA, and brains were left in 4% PFA overnight. Brains were then cryoprotected with 30% sucrose solution and stored in -80C.
Free floating method was used with 40 micron thick coronal sections. For washes and incubations, 24 well-plate was set on a rotating platform at a speed sufficient enough to keep sections moving in the well.
Secondary antibody was incubated for 2 hrs @ RT with the same diluent used for the primary antibody.
Image shows PDGFR-beta positive staining (red) with FITC Dextran perfused blood vessels (green) and DAPI.
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.
Brandon Jun
Verified customer
提交于 Jan 19 2015