Overview

  • Product nameAnti-Hepcidin-25 antibody
    See all Hepcidin-25 primary antibodies
  • Description
    Rabbit polyclonal to Hepcidin-25
  • Tested applicationsSuitable for: WB, IHC-Fr, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide corresponding to Human Hepcidin-25 aa 50 to the C-terminus conjugated to keyhole limpet haemocyanin.
    Database link: P81172
    (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 notesab75883 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

Associated products

Applications

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.

Application Abreviews Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 3 kDa).Can be blocked with Human Hepcidin-25 peptide (ab31875). 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

  • FunctionLiver-produced hormone that constitutes the main circulating regulator of iron absorption and distribution across tissues. Acts by promoting endocytosis and degradation of ferroportin, leading to the retention of iron in iron-exporting cells and decreased flow of iron into plasma. Controls the major flows of iron into plasma: absorption of dietary iron in the intestine, recycling of iron by macrophages, which phagocytose old erythrocytes and other cells, and mobilization of stored iron from hepatocytes (PubMed:22306005).
    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 (PubMed:11113131, PubMed:11034317).
  • Tissue specificityHighest 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 diseaseHemochromatosis 2B
  • Sequence similaritiesBelongs to the hepcidin family.
  • Cellular localizationSecreted.
  • Information by UniProt
  • Database links
  • FormHepcidin 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
    • Hepc20 antibody
    • Hepc25 antibody
    • HEPCIDIN antibody
    • Hepcidin 20 antibody
    • Hepcidin 25 antibody
    • Hepcidin antimicrobial peptide antibody
    • Hepcidin-20 antibody
    • Hepcidin25 antibody
    • HFE2 antibody
    • HFE2B antibody
    • LEAP 1 antibody
    • LEAP-1 antibody
    • LEAP1 antibody
    • Liver expressed antimicrobial peptide antibody
    • Liver-expressed antimicrobial peptide 1 antibody
    • PLTR antibody
    • Putative liver tumor regressor antibody
    see all

Anti-Hepcidin-25 antibody images

  • 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?)
  • 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.
  • 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.
  • 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.

    See Abreview

References for Anti-Hepcidin-25 antibody (ab75883)

This product has been referenced in:
  • Gnana-Prakasam JP  et al. Retinal expression of the serine protease matriptase-2 (Tmprss6) and its role in retinal iron homeostasis. Mol Vis 20:561-74 (2014). Mouse . Read more (PubMed: 24791141) »

See 1 Publication for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Rat Astrocytes)
Specification Rat Astrocytes
Fixative Paraformaldehyde
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 06 2012

Thank you for your reply.

I am still concerned that the resolving conditions are not suitable for such a small protein. I will send on the peptide as an exception. However, I am not sure how it will be used for blocking if a band of the appr...

Read More

Thank you for contacting us.

I am sorry to hear you have been experiencing problems with this antibody. The qulaity of our products is extremely important to us so thanks for getting in touch.

Hepcidin-25 is a very small protein t...

Read More

Thank you very much for your valuable feedback.

We would be very grateful if you could submit an Abreview about this product via the online product datasheet. We always encourage customers to send their results back to us, whether positive o...

Read More

Thank you for your call today and for letting us know about the trouble with this antibody. As we discussed, I am sending a free of charge vial from a new lot on the order ***, which should arrive tomorrow. Please keep me updated about any results usin...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (liver)
Specification liver
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 03 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (293FT cell line)
Specification 293FT cell line
Fixative Formaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Jan 14 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"