Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Adult Oral Epithelia)
Permeabilization
Yes - 0.05% 10% Triton-X
Specification
Adult Oral Epithelia
Blocking step
Gel Block (5% normal donkey serum, 3% BSA, 8% gelatin, and 0.1% Triton X-100 in 1x PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde
Other product details
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: Gel Block (5% normal donkey serum, 3% B
Dilution
1/1000
Secondary antibody
Dilution
1/500
Name
Non-Abcam antibody was used: Rhodamine Red-X (Invitrogen, 1:500)
Host species: Donkey
Clonality: Polyclonal
Conjugation: Rhodamine
Host species: Donkey
Clonality: Polyclonal
Conjugation: Rhodamine
Additional data
Additional Notes
Tissues were microdissected into ice-cold 1x PBS and fixed for 30 min at room temperature (RT) in 4% paraformaldehyde (PFA). After washing with PBS 3x/10min at RT, samples were equilibrated O/N at 4°C in 15% sucrose solution and then mounted in Tissue- Tek optimal cutting temperature (OCT) compound (Electron Microscopy Services). 12µm
sagittal and coronal sections were cut on a Leica CM1950 cryostat onto Fisher SuperFrost Plus slides and stored at -80°C. Samples were dried at 37°C for 30 min before a 1h incubation with gelatin block (5% normal donkey serum, 3% BSA, 8% gelatin, and 0.1% Triton X-100 in 1x PBS). Slides were incubated with primary antibodies (Rt-Ki67 and Rb-EGFR) diluted in gelatin block O/N at 4°C and washed 3x/5min in 1x PBS at RT. Secondary antibodies (Donkey anti-rat Alexa Fluor® 488 and Donkey anti-rabbit Rhodamine Red-X) were also diluted in gelatin block and added to the slide for 2h at RT. DAPI (1:2000) was added to the slide for 5 min at RT samples were mounted in 100µL ProLong Gold (Invitrogen) and covered by glass coverslips (Fisher Scientific).
Abcam response
Thank you for participating in our AbTrial program and your valuable feedback. We are pleased that this antibody worked well for you in IHC-Fr, an application that has not been tested before. We welcome any further feedback and AbReviews from other researchers who have used this antibody for IHC-Fr which will help us assess how well the antibody works in this application.
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.
Bethany Wagner
Verified customer
提交于 Jun 04 2018