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  1. Link

    products/primary-antibodies/hepcidin-25-antibody-ab30760.pdf

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Cardiovascular Blood Serum Proteins
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Anti-Hepcidin-25 antibody (ab30760)

  • Datasheet
  • SDS
Reviews (9)Q&A (2)References (35)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)
  • Western blot - Anti-Hepcidin-25 antibody (ab30760)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)

Key features and details

  • Rabbit polyclonal to Hepcidin-25
  • Suitable for: WB, IHC-P
  • Reacts with: Rat, Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-Hepcidin-25 antibody [EPR18074] (ab187778)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-Hepcidin-25 antibody
    See all Hepcidin-25 primary antibodies
  • Description

    Rabbit polyclonal to Hepcidin-25
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 to the C-terminus of Human Hepcidin-25.

    Read Abcam's proprietary immunogen policy (Peptide available as ab31875.)
  • Positive control

    • WB: Recombinant Hepcidin protein. IHC-P: Human liver tissue.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Serum Proteins
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Cardiovascular
    • Blood
    • Erythrocytes
    • Metabolism
    • Pathways and Processes
    • Cofactors, Vitamins / minerals
    • Vitamins / minerals
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Immunizing Peptide (Blocking)

    • Human Hepcidin-25 peptide (ab31875)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)

Applications

The Abpromise guarantee

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.

Application Abreviews Notes
WB (5)
1/10 - 1/100. Detects a band of approximately 32 kDa (predicted molecular weight: 3 kDa).
IHC-P (1)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
1/10 - 1/100. Detects a band of approximately 32 kDa (predicted molecular weight: 3 kDa).
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

  • 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.
  • Target information above from: UniProt accession P81172 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 57817 Human
    • Entrez Gene: 84506 Mouse
    • Entrez Gene: 84604 Rat
    • Omim: 606464 Human
    • SwissProt: P81172 Human
    • SwissProt: Q9EQ21 Mouse
    • SwissProt: Q99MH3 Rat
    • Unigene: 8821 Human
    • 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
      • 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)

      IHC image of Hepcidin-25 staining in Human normal liver formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab30760, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Western blot - Anti-Hepcidin-25 antibody (ab30760)
      Western blot - Anti-Hepcidin-25 antibody (ab30760)
      Anti-Hepcidin-25 antibody (ab30760) at 1 µg/ml + HAMP Human Full-length Recombinant Protein (Tagged) at 0.1 µg

      Secondary
      Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

      Performed under reducing conditions.

      Predicted band size: 3 kDa
      Observed band size: 32 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      ab30760 was tested against Human HAMP Full-length Recombinant Protein (Tagged) predicted to run at 32kDa.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hepcidin-25 antibody (ab30760)This image is courtesy of Ruma Raha-Chowdhury (Cambridge University)

      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.

    Protocols

    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols
    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (35)

    Publishing research using ab30760? Please let us know so that we can cite the reference in this datasheet.

    ab30760 has been referenced in 35 publications.

    • Raha AA  et al. Interplay of Ferritin Accumulation and Ferroportin Loss in Ageing Brain: Implication for Protein Aggregation in Down Syndrome Dementia, Alzheimer's, and Parkinson's Diseases. Int J Mol Sci 23:N/A (2022). PubMed: 35162984
    • Soliman SSM  et al. Novel Secreted Peptides From Rhizopus arrhizus var. delemar With Immunomodulatory Effects That Enhance Fungal Pathogenesis. Front Microbiol 13:863133 (2022). PubMed: 35387075
    • Hu J  et al. Role of NOD2 and hepcidin in inflammatory periapical periodontitis. BMC Oral Health 22:263 (2022). PubMed: 35764993
    • Chen Q  et al. Electroacupuncture Reduces Cerebral Hemorrhage Injury in Rats by Improving Cerebral Iron Metabolism. Mediators Inflamm 2022:6943438 (2022). PubMed: 36016663
    • Probst S  et al. Role of hepcidin in oxidative stress and cell death of cultured mouse renal collecting duct cells: protection against iron and sensitization to cadmium. Arch Toxicol 95:2719-2735 (2021). PubMed: 34181029
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 11 Abreviews or Q&A

    Western blot abreview for Anti-Hepcidin-25 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Pig Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (4–15% Mini-PROTEAN® TGX™ Precast Protein Gel)
    Loading amount
    20 µg
    Specification
    Liver
    Blocking step
    skimmed milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More
    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.

    DR. Liliana Kiczak

    Verified customer

    Submitted Feb 22 2021

    Western blot abreview for Anti-Hepcidin-25 antibody

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Liver tissue lysate)
    Gel Running Conditions
    Reduced Denaturing (11% SDS-PAGE)
    Loading amount
    30 µg
    Specification
    Liver tissue lysate
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    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 Aug 23 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Hepcidin-25 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Baboon Tissue sections (Adult liver)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM sodium citrate, pH 6.0
    Permeabilization
    No
    Specification
    Adult liver
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C
    Fixative
    Formaldehyde
    Read More
    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.

    MS. Marcia Burch

    Verified customer

    Submitted Mar 22 2016

    Western blot abreview for Anti-Hepcidin-25 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Sample
    Rat Tissue lysate - whole (liver)
    Specification
    liver
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    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 01 2014

    ELISA abreview for Anti-Hepcidin-25 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    ELISA
    Sample
    Mouse Recombinant protein (Mouse cell lysate)
    Specification
    Mouse cell lysate
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
    Type
    Indirect
    Read More
    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 15 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Hepcidin-25 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Rat Cell (Astrocytes culture)
    Specification
    Astrocytes culture
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.3% TritonX in 0.1% PBS
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
    Read More
    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.

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Feb 13 2012

    Western blot abreview for Anti-Hepcidin-25 antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Liver)
    Loading amount
    50 µg
    Specification
    Liver
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    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 Sep 06 2011

    Immunohistochemistry (Frozen sections) abreview for Anti-Hepcidin-25 antibody

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Human Tissue sections (artery)
    Specification
    artery
    Fixative
    Acetone
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 0.5% · Temperature: 20°C
    Read More
    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 Dec 10 2009

    Question

    Dear sir/madam,

    A few months ago I wanted to order the hepcidin antibody ab30760, but unfortunately it was not available then.

    Could you inform me on the status of this antibody now? Is it available again or expected to be available again any time soon?

    Kind regards,

    Read More

    Abcam community

    Verified customer

    Asked on Oct 09 2014

    Answer

    Thank you for your enquiry.

    I am sorry to confirm this antibody is still not available and for the inconvenience. I contacted the laboratories and they confirmed that another batch is currently in production, but due to difficulties with past batches, we cannot yet guarantee this new batch will pass QC and be available for sale.

    We do have another Hepcidin 25 antibody you may want to consider trying:


    Anti-Hepcidin-25 antibody (ab75883)
    Description: Rabbit polyclonal to Hepcidin-25
    Application: ICC/IF, IHC-Fr, WB
    Reactivity: Mouse, Human
    https://www.abcam.com/Hepcidin-25-antibody-ab75883.html (or use the following: https://www.abcam.com/Hepcidin-25-antibody-ab75883.html).

    I am sorry we are not currently able to provide the product you require, but I hope this alternative may be helpful. If you have any questions, please do not hesitate to contact me.

    Read More

    Sam Washer

    Abcam Scientific Support

    Answered on Oct 09 2014

    Question

    LOT NUMBER GR61738-3 ORDER NUMBER xxxxx DESCRIPTION OF THE PROBLEM No signal or weak signal SAMPLE human cellular lysates and secretomes human tissue extracts PRIMARY ANTIBODY Abcam / rat and human / BSA 1%-OVA 3% in TBS-Tween 0.1% / dilution 1μg/mL / incubation O/N 4°C / washed 3 times for 5 up to 10 min DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED positive controls: HepG2 and MCF-7 lysates ANTIBODY STORAGE CONDITIONS aliquoted and stored at -20°C SAMPLE PREPARATION Lysis in PLCLB buffer + protease inhibitors. Samples were heated for 10 min at 70°C before gel loading. AMOUNT OF PROTEIN LOADED 30 μg of total proteins ELECTROPHORESIS/GEL CONDITIONS 1) Reducing gel, 4-12% bis-tris, buffer: MOPS SDS 2) Reducing gel 12% bis-tris, buffer: MES SDS TRANSFER AND BLOCKING CONDITIONS 1) TRANSFER conditions: transfer buffer (INVITROGEN), time: 1h30min 2) BLOCKING conditions: BSA 1%-OVA 3% in TBS-Tween 0.1%, time: 1 up to 2 h. We used both PVDF and nitrocellulose membranes SECONDARY ANTIBODY Sigma / rabbit / BSA 1%-OVA 3% in TBS-Tween 0.1% / dilution 1:4000 / incubation 1h RT / washed 3 times for 10 min HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 3 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? none ADDITIONAL NOTES we saw only bands related to the light and heavy chains of the antibody in a WB and no band in other WB. at the time of the delivery, the antibody was not at proper temperature and the package was a bit damaged and lacking of the datasheet

    Read More

    Abcam community

    Verified customer

    Asked on Dec 02 2011

    Answer

    Thank you for taking time to complete our questionnaire and for contacting us. I am sorry to hear this antibody is not providing satisfactory results and for the delay in getting back to you. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. In order to understand why this antibody might not be working could I please ask you to answer a few additional questions? Has the transfer to the membrane been checked with staining of the membrane with a stain such as Ponceau Red? You say that you are only able to observe light and heavy bands from the antibody, are you performing immunoprecipitation with your sample? In order to obtain stronger staining I would try incubating the with the primary antibody for 1-2 hours at room temperature. I would also suggest just blocking with 3% milk for 1 hour at room temperature and only add 0.1% milk in your antibody diluents. This may help in obtaining a stronger signal from your sample. If however these suggestions do not help then please let me know and we can arrange for a replacement antibody or a refund, whichever you would prefer. In regards to how the antibody was delivered to you, we normally deliver at 4°C with an ice pack and this should maintain the activity of the antibody throughout transport. Could you please describe how you received your antibody and I will look into it. Have you ordered your antibody from one of our distributors or directly from us? I am sorry for the inconvenience this problem is causing. 

    Read More

    Abcam Scientific Support

    Answered on Dec 02 2011

    1-10 of 11 Abreviews or Q&A

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