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Human Hepcidin-25 peptide (ab31875)

Reviews (1)Q&A (9)References (4)

Key features and details

  • Purity: > 70% HPLC
  • Suitable for: Blocking, ELISA

Description

  • Product name

    Human Hepcidin-25 peptide

  • Purity

    > 70 % HPLC.
    70 - 90% by HPLC

  • Accession

    P81172

  • Animal free

    No

  • Nature

    Synthetic

    • Species

      Human

Associated products

Specifications

Our Abpromise guarantee covers the use of ab31875 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    Blocking - Blocking peptide for Anti-Hepcidin-25 antibody (ab30760)

    ELISA

  • Form

    Lyophilized

  • Additional notes

    Storage buffer: 10% 0.5M acetic acid, 90% TK buffer. pH 3.6

    Solutions of ab31875, diluted 10 times, in TBST (80%) and PBS (10%) have a pH of 5.1

  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.

    Information available upon request.

  • Reconstitution
    Reconstitute in 10% 0.5M acetic acid, 90% TK buffer. If the peptide doesn’t dissolve try an organic solvent like DMSO, then dilute using water or buffer. Gentle warming and sonication can effectively aid peptide solubilisation

General Info

  • Alternative names

    • Hamp
    • HEPC
    • HEPC_HUMAN
    • Hepc20
    • Hepc25
    • HEPCIDIN
    • Hepcidin 20
    • Hepcidin 25
    • Hepcidin antimicrobial peptide
    • Hepcidin-20
    • Hepcidin25
    • HFE2
    • HFE2B
    • LEAP 1
    • LEAP-1
    • LEAP1
    • Liver expressed antimicrobial peptide
    • Liver-expressed antimicrobial peptide 1
    • PLTR
    • Putative liver tumor regressor
    see all

  • Function

    Liver-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 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

    Hemochromatosis 2B

  • Sequence similarities

    Belongs to the hepcidin family.

  • Cellular localization

    Secreted.
  • Information by UniProt

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (4)

Publishing research using ab31875? Please let us know so that we can cite the reference in this datasheet.

ab31875 has been referenced in 4 publications.

  • Zhou X  et al. Tertiary lymphoid structure stratifies glioma into three distinct tumor subtypes. Aging (Albany NY) 13:26063-26094 (2021). PubMed: 34954691
  • Xiao L  et al. Macrophage iron retention aggravates atherosclerosis: Evidence for the role of autocrine formation of hepcidin in plaque macrophages. Biochim Biophys Acta Mol Cell Biol Lipids 1865:158531 (2020). PubMed: 31666189
  • Xiong XY  et al. Toll-Like Receptor 4/MyD88-Mediated Signaling of Hepcidin Expression Causing Brain Iron Accumulation, Oxidative Injury, and Cognitive Impairment After Intracerebral Hemorrhage. Circulation 134:1025-1038 (2016). PubMed: 27576776
  • McCarthy RC & Kosman DJ Glial cell ceruloplasmin and hepcidin differentially regulate iron efflux from brain microvascular endothelial cells. PLoS One 9:e89003 (2014). PubMed: 24533165

Customer reviews and Q&As

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1-10 of 10 Abreviews or Q&A

ELISA Abreview for Hepcidin-25 peptide

 Good
Abreviews
abreview image
Application
ELISA
Review text: Used in conjunction with ab30760 at does dependent concentration as a positive control.
Sample: Mouse Recombinant protein (Mouse Primary neurons)
Specification: Mouse Primary neurons
Blocking step: BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 37°C
Type: Direct

Other product details
Incubation time: 1 hour(s) and 0 minute(s) · Temperature: 37°C · Diluent: ([2.93g Sodium Bicarbonate (NaHCO3), 1.5

Primary antibody (in addition to 'Hepcidin-25 peptide')
Dilution: 1/5000

Secondary antibody
Name: Non-Abcam antibody was used: Dako goat anti rabbit HRP antibody
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution: 1/50000
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 09 2012

Question

We're planning to test Hepcidin-25 in an animal basic, could you tell us what kind of solution the peptide is in?

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Answer

Thank you for contacting Abcam regarding ab31875.


The buffer contains the following:


30 mM HEPES
150 mM NaCl
2 mM EDTA
0.001% Tween 20
At pH 6.75


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Question

Je vous remercie d’avoir traité si rapidement ma requête. J’accepte votre proposition à savoir un avoir/remboursement.
Bien cordialement

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Answer

Merci pour la confirmation.

Comme convenu, j'ai demandé à notre service comptabilité de mettre en place un avoir référence CN.

Votre référence CN est *******.

Voici les manières dont vous pouvez utiliser cette référence CN:
(1) la faire valoir contre la facture originale si celle-ci n'as pas encore été payée
(2) la faire valoir contre une prochaine facture
(3) obtenir un remboursement de la somme entière si vous n'avez pas de factures actuellement en cours avec Abcam

Si vous souhaitez recevoir un remboursement au lieu d'un avoir, veuillez demander à votre service de gestion de prendre contact avec notre département comptabilité afin que nous puissions recueillir les informations nécessaires pour cette restitution. Notre service comptabilité peut être contacté par courriel à l'adresse mailto:creditcontrol@abcam.com ou par téléphone en utilisant le lien « Contactez-nous » dans le coin en haut à droite de notre site internet. Veuillez communiquer votre numéro d'avoir lorsque vous contactez notre service comptabilité.

Vous recevrez également la référence complète de la note de crédit par courriel ou par voie postale; cette référence commençant par les lettres CGB.

J'espère que cette expérience ne vous empêchera pas de commander à nouveau l'un de nos produits.

L'équipe technique reste à votre disposition pour plus de conseils.

Bonne continuation dans votre recherche.

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Question

Product code: 31875
Lot number: GR76589-1

Inquiry: I'm working on the development of ELISA Hepcidin.
Antibodies anti-Hepcidin generated at Diaclone and an other one generated by a collaborator well recognize hepcidin purchased from a concurrent but do not recognize yours.
I tested it with a sandwich ELISA and with a direct ELISA, without success.

Please see above our experimental conditions:
- Direct ELISA : coating : Hepcidin 500ng/ml (100µl/well) in PBS O/N +4°C Wash blocking: 250µl/well BSA 5% 2H RT Wash primary antibody : 10µg/ml diluted 10 fold on 7 points (BSA1%) 1H RT Wash secondary antibody : goat anti-mouse IG1 HRP, 1H RT Wash Substrate: TMB 100µl/well 15' RT Stop with 100µl/well H2SO4 -Sandwich ELISA coating: capture antibody 10µg/ml(100µl/well) in PBS O/N +4°C wash blocking: 250µl/well BSA 5% 2H RT Wash Hepcidin standard : 1µg/ml diluted 2 fold on 7 points (BSA1%) 2H RT Wash biotinylated detection antibody : 1µg/ml (100µl/well) (BSA 20%) 1H RT Wash Streptavidin HRP : 100µl/well 30' RT wash Substrate: TMB 100µl/well 15' RT Stop with 100µl/well H2SO4 See in attachment data obtained.

Do you already have a similar request with this product?

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Answer

Merci de nous avoir contactés.

Je suis désolé que le produit ne fonctionne pas.

L'explication est très probablement la suivante: la longueur de ce peptide n'est que de 25 acides aminés (aa 60-84 de l'hépicine humaine). Ce peptide sert d'immunogène de l'anticorps ab75883.

J'ai parcouru la fiche technique qui, j'en suis désolé, contient une information qui porte à confusion , à savoir la longueur de la protéine (et non du peptide) qui est de 84 acides aminés.

J'ai demandé à ce que cette information soit reformulée ou supprimée de la fiche technique.

Nous n'avons pas à notre catalogue l'hépicine entière. Je veux bien néanmoins procéder à l'édition d'un avoir / remboursement en compensation de cette confusion.

Merci de me confirmer que cette résolution vous convient.

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Question

I would like to know in which buffer I should dissolve the Hepcidin-25 peptide (ab31875). Is water ok? Thank you very much.

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Answer

Thank you for contacting us.

The peptide should already be in solution. Is that not the case? You should have 100ul. Be sure to centrifuge the tube to pulll down any solution trapped in the threads of the tube.

If you want to further dilute the stock solution, distilled water will be fine.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

Do you have information of the reactivity of "Hepcidin-25 peptide (ab31875)" on mouse cells? Or is it possible to obtain a test-sample to test the reactivity?

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Answer

Thank you for contacting us.

The product ab31875 is a synthetic 25 amino acid peptide that is not, to our knowledge, active. This peptide has been used to produce the antibody ab75883, which is validated for use on mouse and human samples in WB, IHC-Fr, and ICC/IF. I hope this helps, please let me know if you have any other questions.

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Question

We appreciated the advice we have obteined from Abcam, but I communicate that the use, in our experiments, of 12% gels with MES SDS buffer is equivalent to having done running the proteins at in a gel of approximately 18% polyacrylamide, so we are confident that the protein of our interest has not been lost.

Please let me know if you can send the the immunizing peptide for blocking experiments.

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Answer

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 appropriate size is not detected.

Please confirm the order number and date of shipping so that I can send on the peptide. Looking forward to your reply.

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Question

Thanks very much for your detailed response, that’s helpful. Just a couple of further questions/comments: when you state that the peptide sequence corresponds to aas 60-84, are you referring to ab31876 or ab31875 or both? There is evidence in the literature that pro-hepcidin is found in the blood as well as located intracellularly, and pre-pro-hepcidin can also be secreted from cells. I do think it might be helpful for customers to know that the ‘hepcidin-25’ antibodies are likely to recognise other forms of hepcidin (since it can be important to distinguish between active hepcidin and inactive precursors), and therefore it would be good to provide this information on the data sheet.  I hope to place an order for one of the hepcidin peptides and some anti-hepcidin antibodies soon. Best wishes,

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Answer

Thanks you for your email. Dr. Tanya is away this week so I am looking after her inbox. The peptide sequence for ab31876 corresponds to amino acid XXXX and peptide sequence for ab31875 corresponds to XXXX amino acids. The antibody cross reactivity always depends on the immunogen sequences, so customers will automatically know from the datasheet e.g. which part of the immunogen the antibody is specific to and in which part of the processed protein, the immunogen lies. This is very obvious information so we do not think we need to publish this information. However we are always happy to answer any inquiry we receive regarding there products. I hope this will be helpful. Should you any other please does not hesitate to contact us.

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Question

Hi, I have a query about this product (hepcidin-25 peptide). I don't understand why this product is termed 'hepcidin-25' when it is actually full length pre-pro-hepcidin rather than the 25 amino acid active form of hepcidin. It's very misleading, can you explain what you mean by 'hepcidin-25' here? I'm also confused about the 'hepcidin-25' antibodies you have - are these specific to the 25 aa form or the 84 aa protein? Many thanks for your help,

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Answer

Thank you for your enquiry and your interest in our products. I have conducted a quick blast and the peptide sequence picks up both hepcidin and hepcidin 25. The peptide sequence corresponds to aas 60-84 of the full length protein. The antibody recognizes the processed hepcidin 25 (60-84 aa) which will be found at low concentrations in the blood (presumably) and it will also detect the full length pre-pro protein which would be found in the cell – hence easier to run on a Western blot. Therefore it would be very difficult to get an antibody specific for the processed protein. I hope this helps and if I can assist further, please do not hesitate to contact me.

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Question

Can you please tell me how this peptide was fabricated and purified?

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Answer

Thank you for contacting us. This peptide was synthesised on solid phase, using Fmoc-protected amino acids. Amino acid side-chains are protected. The full length peptide is cleaved from the resin and fully deprotected using 95% TFA and scavenger mix to remove protecting group by-products. The crude peptide is purified by reverse phase HPLC, using aqueous acetonitrile with 0.1% gradient. The peptide pool is then lyophilized, and is supplied in fully reduced form (ie no disulfide bridges), and TFA salt. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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