Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Primary Astrocytes culture)
Specification
Primary Astrocytes culture
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10%
Other product details
Dilution
1/500
Incubation time
4 hour(s) and 0 minute(s) · Temperature: 24°C
Secondary antibody
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 (Ab39742): Cultured astrocytes were stained with polyclonal anti-GPCR GPR30. 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 (Ab66048, 1/500 dilution) 4 hours at 24°C. The secondary antibody (green) was Alexa Fluor® 488 donkey anti-rabbit IgG (H+L) used at a 1/1000 dilution and monoclonal antiGFAP-568for 1h. Hoechst was used to stain the cell nuclei (blue) at a concentration of 1.43µM. GPCR expressed in selected astrocytes.
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
提交于 Feb 13 2012