Immunocytochemistry/ Immunofluorescence abreview for Anti-Hepcidin-25 antibody

Excellent
Abreviews
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Astrocytes culture)
Specification
Astrocytes culture
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Other product details

Dilution
1/100
Incubation time
24 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 0.3% TritonX with 0.1x PBS- 10% Donkeys

Secondary antibody

Name
Non-Abcam antibody was used: anti rabbit Alexa Fluro 488
Host species: Donkey
Clonality: Polyclonal
Conjugation: Alexa Fluor® 488
Dilution
1/1000

Additional data

Additional Notes
ICC/IF image of (ab30760): Cultured astrocytes were stained with polyclonal anti-hepcidin. 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 (ab30760, 1/100 dilution) overnight at +4°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. Hepcidin expression visible in perinuclear mainly in the lysosomes.

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 13 2012

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