Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections (30 um free floating))
Specification
Brain sections (30 um free floating)
Fixative
Paraformaldehyde
Antigen retrieval step
None
Permeabilization
No
Other product details
Dilution
1/3000
Incubation time
18 hour(s) and 0 minute(s) · Temperature: 20°C · Diluent: PBS triton 0.3%
Secondary antibody
Name
Non-Abcam antibody was used: Goat anti-rabbit alexa Fluor 488
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
ab16550 produced a nice cytoplasmic staining in some neurons of the central nervous system (in naïve animals), such as some cortical and some hypothalamic neurons. The staining is observed in the soma and processes of these neurons.
The staining is quenched by pre-incubation with peptide against phospho S847 (ab16981), but is NOT quenched using the control peptide (ab57047), suggesting that the staining obtained with the antibody is specific of the phosphorylated form S847.
ab16650 was used at 1/3000 incubated overnight at room temperature. Secondary antibody used was anti-rabbit Alexa Fluor 488 at 1/1000 for 2 hours. Animals were perfused-fixed with paraformaldehyde 4%. Tissues were post-fixed overnight in the same fixative and then cryoprotected in 20% sucrose overnight. Following embedding in OCT and freezing, tissues were cut (30 um thickness) and immunostained using the 'free floating’ technique.
The staining is quenched by pre-incubation with peptide against phospho S847 (ab16981), but is NOT quenched using the control peptide (ab57047), suggesting that the staining obtained with the antibody is specific of the phosphorylated form S847.
ab16650 was used at 1/3000 incubated overnight at room temperature. Secondary antibody used was anti-rabbit Alexa Fluor 488 at 1/1000 for 2 hours. Animals were perfused-fixed with paraformaldehyde 4%. Tissues were post-fixed overnight in the same fixative and then cryoprotected in 20% sucrose overnight. Following embedding in OCT and freezing, tissues were cut (30 um thickness) and immunostained using the 'free floating’ technique.
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.
Dr. Sophie Pezet
Verified customer
提交于 Jan 17 2008