Anti-Hepcidin-25 antibody (ab30760)

Immunohistochemistry (PFA fixed) - Hepcidin-25 antibody (ab30760)

Hepcidin-25 expression in Rat liver cells using Rabbit polyclonal to Hepcidin-25 (ab30760).Immunohistochemistry was performed on PFA fixed cells, ab30760 was used at 1/1000 incubated overnight at RT. Haemotoxylin is used as a nuclear counterstain in this image.

Ruma Raha-Chowdhury, University Of Cambridge, United Kingdom

Western blot - Hepcidin-25 antibody (ab30760)

Anti-Hepcidin-25 antibody (ab30760) at 1 µg/ml + HAMP Human Full-length Recombinant Protein (Tagged) at 0.1 µg

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 3 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)


Exposure time : 3 minutes

ab30760 was tested against Human HAMP Full-length Recombinant Protein (Tagged) predicted to run at 32kDa.

Immunohistochemistry (Frozen sections) - Hepcidin-25 antibody (ab30760)

ab30760 at a 1/200 dilution staining Hepcidin-25 in human artery tissue sections by Immunohistochemistry (frozen sections) incubated for 24 hours at 20°C. Acetone fixed. Blocked with 0.5% BSA for 10 minutes at 20°C. Secondary used at 7µg/ml Goat anti-rabbit IgG (H+L) conjugated to biotin.

Immunoperoxydase system and AEC were used to develop the color.

This image is courtesy of an anonymous abreview.

Immunocytochemistry/ Immunofluorescence - Anti-Hepcidin-25 antibody (ab30760)

ab30760 staining cultured rat astrocytes by ICC/IF. The cultured cells were fixed with 4% paraformaldehyde for 5 minutes and blocked with 10% donkey serum in 0.1% PBS-0.3% TritonX for 30 minutes at 24°C. The cultured cells were then stained with ab30760 at 1/100 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 24h at 4°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue. Hepcidin expression visible perinuclearly,  mainly in the lysosomes.

This image is courtesy of an Abreview submitted by Ruma Raha-Chowdhury