Recombinant
RabMAb

Recombinant Anti-Hepcidin + Hepcidin-2 antibody [EPR18937] (ab190775)

Overview

  • Product name

    Anti-Hepcidin + Hepcidin-2 antibody [EPR18937]
    See all Hepcidin + Hepcidin-2 primary antibodies
  • Description

    Rabbit monoclonal [EPR18937] to Hepcidin + Hepcidin-2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse Hepcidin + Hepcidin-2 aa 1 to the C-terminus. The exact sequence is proprietary. Hepcidin-2 = SwissProt: Q80T19
    Database link: Q9EQ21

  • Positive control

    • WB: Mouse Hepcidin recombinant fragment protein; Mouse Hepcidin-2 recombinant fragment protein; Mouse liver lysate. IHC-P: Mouse liver tissue. IHC-F: Mouse liver tissue.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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 long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18937
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab190775 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
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/200. Detects a band of approximately 9 kDa (predicted molecular weight: 9 kDa).
IHC-Fr 1/100.

Target

  • Relevance

    Hepcidin: Q9EQ21: 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. Hepcidin-2: Q80T19: Seems to act as a signaling molecule involved in the maintenance of iron homeostasis.
  • Database links

  • Alternative names

    • Hamp antibody
    • Hamp1 antibody
    • Hamp2 antibody
    • Hepc antibody
    • Hepc1 antibody
    • Hepc2 antibody
    see all

Images

  • Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Cytoplasmic staining on hepatocytes of Mouse liver was observed.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

  • All lanes : Anti-Hepcidin + Hepcidin-2 antibody [EPR18937] (ab190775) at 1/200 dilution

    Lane 1 : Mouse liver lysate
    Lane 2 : Mouse brain lysate
    Lane 3 : Mouse heart lysate
    Lane 4 : Mouse kidney lysate
    Lane 5 : Mouse spleen lysate
    Lane 6 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 7 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 9 kDa
    Observed band size: 9 kDa


    Exposure time: 20 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Highly expressed in the liver and to a much lesser extent in the heart (PMID:11113132 and PMID:12729891)

  • All lanes : Anti-Hepcidin + Hepcidin-2 antibody [EPR18937] (ab190775) at 1/5000 dilution

    Lane 1 : Mouse Hepcidin recombinant fragment protein
    Lane 2 : Mouse Hepcidin-2 recombinant fragment protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 9 kDa
    Observed band size: 32 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time:

    Lane 1: 1 second.
    Lane 2: 2 seconds.

    The mouse recombinant fragment proteins contain aa24-83 with a GST-Tag and were made in-house.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized frozen section of Mouse liver labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)secondary antibody at 1/1000 dilution (green).

    Confocal image showing cytoplasmic staining on hepatocytes of Mouse liver.

    Counter stained with DAPI.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

  • Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Mouse kidney. Counter stained with Hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Mouse spleen. Counter stained with Hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Rat liver. This could be because the ab does not XR with Rat hepcidin (77% identity between mouse and rat hapcidin sequences).

    Counter stained with Hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Hepcidin + Hepcidin-2 with ab190775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative staining on Human colon cancer. Counter stained with Hematoxylin.

References

This product has been referenced in:

  • Fader KA  et al. Convergence of hepcidin deficiency, systemic iron overloading, heme accumulation, and REV-ERBa/ß activation in aryl hydrocarbon receptor-elicited hepatotoxicity. Toxicol Appl Pharmacol 321:1-17 (2017). Read more (PubMed: 28213091) »
See 1 Publication for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Serum (Human serum and LPS treated rat serum)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Treatment
rat treated with 5mg/kg LPS and then serum collected aftrer 4 hours
Specification
Human serum and LPS treated rat serum
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Dec 08 2015

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