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Products:Cardiovascular >> Blood >> Serum Proteins
Overview
Product name
Anti-Hepcidin-25 antibody
See all Hepcidin-25 products (8) ...
Description
Rabbit polyclonal to Hepcidin-25
Tested applications
WB, IHC-Fr, ICC/IFmore details
Cross reactivity
Reacts with
Mouse, Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Hepcidin-25.
(Peptide available as ab31875.)
Positive control
This antibody gave a positive signal against Human HAMP Full-length Recombinant Protein (Tagged) (this explains why the observed band 30kDa is so much larger than the expected 2.8kDa band)
General notes
ab75883 is raised using the same immunogen as ab30760. Batches sold under ab75883 have passed QC testing in Western Blot (Recombinant only) and ICC/IF. In our hands, this product did not work in IHC-P. For further information please contact our Customer Services team.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Cardiovascular >> Blood >> Erythrocytes
Signal Transduction >> Metabolism >> Vitamins / Minerals
Cardiovascular >> Blood >> Serum Proteins
Applications
Show applications key
Our Abpromise guarantee covers the use of ab75883 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 1 µg/mlDetects a band of approximately 32 kDa (predicted molecular weight: 3 kDa).(No data on endogenous protein, only tested in recombinant protein. )
IHC-Fr: 1/250
ICC/IF: Use a concentration of 1 - 5 µg/ml.
Target
Function
Seems to act as a signaling molecule involved in the maintenance of iron homeostasis. Seems to be required in conjunction with HFE to regulate both intestinal iron absorption and iron storage in macrophages.
Has strong antimicrobial activity against E.coli ML35P N.cinerea and weaker against S.epidermidis, S.aureus and group b streptococcus bacteria. Active against the fungus C.albicans. No activity against P.aeruginosa.
Tissue specificity
Highest expression in liver and to a lesser extent in heart and brain. Low levels in lung, tonsils, salivary gland, trachea, prostate gland, adrenal gland and thyroid gland. Secreted into the urine.
Involvement in disease
Defects in HAMP are the cause of hemochromatosis type 2B (HFE2B) [MIM:613313]; also known as juvenile hemochromatosis (JH). HFE2B is a disorder of iron metabolism with excess deposition of iron in the tissues, bronze skin pigmentation, hepatic cirrhosis, arthropathy and diabetes. The most common symptoms of hemochromatosis type 2 at presentation are hypogonadism and cardiomyopathy.
Sequence similarities
Belongs to the hepcidin family.
Cellular localization
Secreted.
Target information above from: UniProt accessionP81172
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Form
Hepcidin produces a propeptide of 84 amino acids that undergoes enzymatic cleavage into mature peptides of 20, 22, and 25 amino acids. Active peptides are rich in cysteines that form intramolecular bonds and stabilize the beta-sheet structure
Alternative names
- HAMP antibody
- HEPC antibody
- HEPC_HUMAN antibody
see all
Database links
- Entrez Gene: 57817 Human
- Entrez Gene: 84506 Mouse
- Omim: 606464 Human
- SwissProt: P81172 Human
- SwissProt: Q9EQ21 Mouse
- Unigene: 8821 Human
Anti-Hepcidin-25 antibody images:
Western blot - Hepcidin-25 antibody (ab75883)
Anti-Hepcidin-25 antibody (ab75883) at 1 µg/ml + Human HAMP Full-length Recombinant Protein (Tagged) at 0.1 µg
Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Predicted band size : 3 kDa
Observed band size : 32 kDa (why is the actual band size different from the predicted?)
ab75883 was tested against Human HAMP Full-length Recombinant Protein (Tagged) predicted to run at 32kDa.
Immunocytochemistry/ Immunofluorescence - Hepcidin-25 antibody (ab75883)
ICC/IF image of ab75883 stained HeLa 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 (ab75883, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabit IgG (H+L) used at a 1/1000 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). This antibody also gave a positive result in 4% PFA fixed (10 min) HEK293, HepG2, and MCF-7 cells at 1µg/ml.
Immunocytochemistry/ Immunofluorescence - Hepcidin-25 antibody (ab75883)
ICC/IF image of ab75883 stained HepG2 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 (ab75883, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 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). This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, MCF-7 cells at 5µg/ml, and in 100% Methanol fixed (5 min) Hek293, HepG2, and MCF-7 cells at 5µg/ml.
Immunohistochemistry (Frozen sections) - Hepcidin-25 antibody (ab75883)
ab75883 staining Hepcidin-25 in Mouse liver tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with formaldehyde, permeabilized with 0.1% Triton X 100 and blocked with 5% serum for 1 hour at 25°C. Samples were incubated with primary antibody (1/250 in PBS Tween) for 12 hours at 4°C. An Alexa Fluor®594-conjugated goat anti-rabbit IgG polyclonal (1/250) was used as the secondary antibody. Nuclei were stained with DAPI.
This image is courtesy of an anonymous Abreview
References for Anti-Hepcidin-25 antibody (ab75883)
ab75883 has not yet been referenced specifically in any publications.
Publishing research using ab75883? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"