Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 24°C
Sample
Mouse Tissue sections (Spleen)
Specification
Spleen
Permeabilization
Yes - Triton X-100 (0.3%)
Fixative
Paraformaldehyde
Other product details
Incubation time
16 hour(s) and 0 minute(s) · Temperature: 4°C
Dilution
1/100
Secondary antibody
Name
Non-Abcam antibody was used: Alexa Fluor 555 Goat anti-rabbit IgG (H+L)
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 555
Host species: Goat
Clonality: Polyclonal
Conjugation: Alexa Fluor 555
Dilution
1/250
Additional data
Additional Notes
The tissues were taken out of the animal and immediately submerged into a mild fixation buffer (PLP) containing 1% paraformaldehyde. The tissues were placed into 30% sucrose for ~ 6 hours before freezing them into OCT tissue compound. For this experiment, splenic tissue sections of a wt mouse were stained for DAPI (Blue), rat anti-CD3 from a different company (green), and rabbit anti-CD3 from abcam (ab5690)(red). Overlap of the two CD3 antibodies is shown in yellow.
Left panel: Secondary antibody only;
Right panel: DAPI (blue), CD3 (green), CD3 (ab5690 - red), CD3 overlap (yellow).
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.
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提交于 Feb 10 2015