Application
Immunohistochemistry (Frozen sections)
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Sample
Mouse Tissue sections (spleen)
Specification
spleen
Permeabilization
No
Fixative
Paraformaldehyde
Other product details
Incubation time
2 hour(s) and 0 minute(s) · Temperature: 24°C · Diluent: PBS
Dilution
1/50
Secondary antibody
Name
Non-Abcam antibody was used: Invitrogen Goat anti Rabbit AF488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000
Additional data
Additional Notes
10um frozen section. Tissue fixed for 1h in 4%PFA, then 20% sucrose for 1h both at RT prior to embedding.
Wash 3x3min in PBS.
Block 10% goat serum for 30min at RT.
Primary Ab diluted in PBS and incubated overnight at 4deg, also works by incubating for 2 hours at RT.
Wash 3x3min in PBS.
Secondary - goat anti-rabbit AF488 (green) 1/1000 in PBS for 1h at RT.
Wash 3x3min in PBS
Counterstained with Hoechst 33258 (blue) for 10mins at RT.
Wash 3min in dH2O
Mounted in Invitrogen ProLong gold and kept at 4deg until imaged.
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提交于 Apr 17 2014