Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Astrocytes culture)
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Specification
Astrocytes culture
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Other product details
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 24°C
Dilution
1/1000
Secondary antibody
Dilution
1/1000
Name
Non-Abcam antibody was used: anti rabbit Alexa Fluro 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Additional data
Additional Notes
ICC/IF image of (ab16640): Cultured astrocytes were stained with polyclonal anti-Sortilin. The cells were 4% formaldehyde fixed (5min) and then incubated in 10% normal donkeys serum in 0.1% PBS-0.3% TritonX for 30 minutes to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16640, 1/1000 dilution) four hours at +24°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Hoechst was used to stain the cell nuclei (blue) at a concentration of 1.43µM. Sortilin expressed in the cytosol mainly in trans-golgi compartments.
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. Ruma Raha-Chowdhury
Verified customer
投稿 Feb 13 2012