Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain)
Permeabilization
Yes - 3% Triton X100
Specification
Brain
Blocking step
Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 0.5µg/mL · Temperature: 23°C
Fixative
Paraformaldehyde
Other product details
Incubation time
1 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 10 mg/ml BSA in PBS
Concentration
100 µg/ml
Secondary antibody
Name
Abcam antibody:Goat Anti-Rabbit IgG Fc (Alexa Fluor® 594)
Additional data
Additional Notes
The experiment was performed in rat brain tissue fixed with 4% paraformaldehyde, 30 micrometers thick, through the free-floating method. Tissues were washed with PBS (3 times, 5 min each wash) and then incubated in 2%BSA / 10% normal goat serum /10% Triton X100 in PBS for 1h to permeabilize the sample and to block non-specific protein interactions. Tissues were then incubated with the antibody (ab41174, 0.5 microgram/mL) overnight at room temperature. The secondary antibody (red) was Alexa Fluor® 594 (ab150092) goat anti-rabbit IgG used at a 1/1000 dilution for 1h. DAPI was used to stain the cell nuclei (blue).
Note to Abcam: Image was captured by a widefield fluorescence. We are planning to repeat this experiment and view it in a confocal microscope, and we are willing to submit images that we can capture in future experiments.
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 Jun 15 2015