The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 5 µg/ml.
1/250. Detects a band of approximately 130 kDa (predicted molecular weight: 130 kDa).
FunctionMay function as a ferroxidase for ferrous (II) to ferric ion (III) conversion and may be involved in copper transport and homeostasis. Implicated in iron homeostasis and may mediate iron efflux associated to ferroportin 1.
Tissue specificityDetected in breast, colon, bone trabecular cells and fibroblasts.
Sequence similaritiesBelongs to the multicopper oxidase family. Contains 6 plastocyanin-like domains.
Immunocytochemistry/ Immunofluorescence - Anti-Hephaestin antibody (ab108003)This image is courtesy of an abreview submitted by Dr. Ruma Raha-Chowdhury, Cambridge University, United Kingdom.
ICC/IF image of Hephaestin staining on rat hippocampal culture using ab108003 (1:200). The cells were fixed with paraformaldehyde and permeabilized using 0.1% TritonX in 0.1% PBS. The cells were blocked using 10 % donkey serum for 1 hour at 24°C. ab108003 was diluted 1:200 using 0.1% TritonX with 0.1x PBS- 10% Donkeys and incubated with the slides at 4°C for 24 hours. The secondary antibody used was donkey polyclonal against rabbit IgG conjugated to Alexa Fluor 488 (1:1000)
ICC/IF image of ab108003 stained MCF7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab108003, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and HepG2 cells at 5µg/ml.
References for Anti-Hephaestin antibody (ab108003)
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