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Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Hepcidin-25.
(Peptide available as ab31875.)
Our Abpromise guarantee covers the use of ab30760 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/10 - 1/100. Detects a band of approximately 32 kDa (predicted molecular weight: 3 kDa).
Current batches of ab30760 gave a positive signal in LPS treated serum in WB at a dilution ranging from 1/10 and 1/100. We recommend customers to determine the optimal dilution for their experiments.
|IHC-P||Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ELISA||Use at an assay dependent concentration.|
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.
This image is courtesy of Ruma Raha-Chowdhury (Cambridge University)
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.
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.
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