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RecombinantRabMAb

Recombinant Anti-HRG antibody [EPR24184-52] (ab281563)

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Western blot - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Immunoprecipitation - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
  • Anti-HRG antibody [EPR24184-52] (ab281563)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR24184-52] to HRG
  • Suitable for: IHC-P, IP, WB
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-HRG antibody [EPR24184-52]
    See all HRG primary antibodies

  • Description

    Rabbit monoclonal [EPR24184-52] to HRG

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, IP, WBmore details
    Unsuitable for: Flow Cyt (Intra),ICC/IF or IHC-Fr

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human plasma and liver tissue lysate. IHC-P: Human liver and colon tissue lysate. IP: Human liver tissue lysate.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

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: 59.94% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR24184-52

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab281563 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
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Predicted molecular weight: 60 kDa.
Notes
IHC-P
1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Predicted molecular weight: 60 kDa.
Application notes
Is unsuitable for Flow Cyt (Intra),ICC/IF or IHC-Fr.

Target

  • Function

    Plasma glycoprotein that binds a number of ligands such as heme, heparin, heparan sulfate, thrombospondin, plasminogen, and divalent metal ions. Binds heparin and heparin/glycosaminoglycans in a zinc-dependent manner. Binds heparan sulfate on the surface of liver, lung, kidney and heart endothelial cells. Binds to N-sulfated polysaccharide chains on the surface of liver endothelial cells. Inhibits rosette formation. Acts as an adapter protein and is implicated in regulating many processes such as immune complex and pathogen clearance, cell chemotaxis, cell adhesion, angiogenesis, coagulation and fibrinolysis. Mediates clearance of necrotic cells through enhancing the phagocytosis of necrotic cells in a heparan sulfate-dependent pathway. This process can be regulated by the presence of certain HRG ligands such as heparin and zinc ions. Binds to IgG subclasses of immunoglobins containing kappa and lambda light chains with different affinities regulating their clearance and inhibiting the formation of insoluble immune complexes. Tethers plasminogen to the cell surface. Binds T-cells and alters the cell morphology. Modulates angiogenesis by blocking the CD6-mediated antiangiongenic effect of thrombospondins, THBS1 and THBS2. Acts as a regulator of the vascular endothelial growth factor (VEGF) signaling pathway; inhibits endothelial cell motility by reducing VEGF-induced complex formation between PXN/paxillin and ILK/integrin-linked protein kinase and by promoting inhibition of VEGF-induced tyrosine phosphorylation of focal adhesion kinases and alpha-actinins in endothelial cells. Also plays a role in the regulation of tumor angiogenesis and tumor immune surveillance. Normalizes tumor vessels and promotes antitumor immunity by polarizing tumor-associated macrophages, leading to decreased tumor growth and metastasis.

  • Tissue specificity

    Expressed in macrophages and in malignant cells. Expressed by the liver and secreted in plasma (at protein level).

  • Involvement in disease

    Thrombophilia due to histidine-rich glycoprotein deficiency

  • Sequence similarities

    Contains 2 cystatin domains.

  • Domain

    The His/Pro-rich (HRR) region contains approximately 12 tandem internal repeats of the 5-residue G[H/P][H/P]PH consensus sequence. HRR binds heparan sulfate and possesses antiangiogenic, antibacterial and antifungal properties through binding Candida cells, and preferentially lysing the ergosterol-containing liposomes at low pH. The tandem repeats also bind divalent metal ions and heme.
    The cystatin domains can also bind heparan sulfate. Binding is enhanced in the presence of zinc ions.

  • Post-translational
    modifications

    Proteolytic cleavage produces several HRG fragments which are mostly disulfide-linked and, therefore, not released. Cleavage by plasmin is inhibited in the presence of heparin, zinc ions or in an acidic environment. Cleavage reduces binding of HRG to heparan sulfate, but enhances the ability of HRG to bind and tether plasminogen to the cell surface. On platelet activation, releases a 33 kDa antiangiogenic peptide which encompasses the HRR. Also cleaved in the C-terminal by plasmin.
    N-glycosylated.

  • Cellular localization

    Secreted.
  • Information by UniProt

  • Database links

    • Alternative names

      • DKFZp779H1622 antibody
      • Histidine proline rich glycoprotein antibody
      • Histidine rich glycoprotein antibody
      • Histidine-proline-rich glycoprotein antibody
      • Histidine-rich glycoprotein antibody
      • HPRG antibody
      • HRG antibody
      • HRG_HUMAN antibody
      • HRGP antibody
      • Thrombophilia due to elevated HRG antibody
      see all

    Images

    • Western blot - Anti-HRG antibody [EPR24184-52] (ab281563)
      Western blot - Anti-HRG antibody [EPR24184-52] (ab281563)
      All lanes : Anti-HRG antibody [EPR24184-52] (ab281563) at 1/1000 dilution

      Lane 1 : Human plasma at 20 µl
      Lane 2 : Human liver tissue lysate at 20 µg
      Lane 3 : Human pancreas tissue lysate at 20 µg

      Secondary
      Lane 1 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
      Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 60 kDa



      The MW and expression pattern is consistent to previous referecence (PMID:14744774, PMID:25363753, PMID:19712047)
      Low expression or negative: human pancreas (PMID:25363753)

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: Lane 1: 3.25 seconds; 

      Lane 2-3: 3 minutes.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)

      Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling HRG with ab281563 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human liver. (PMID: 21215706) is observed. The section was incubated with ab281563 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

      Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)

      Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling HRG with ab281563 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the plasma and stroma cells in human colon (PMID: 25243896) is observed. The section was incubated with ab281563 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

      Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunoprecipitation - Anti-HRG antibody [EPR24184-52] (ab281563)
      Immunoprecipitation - Anti-HRG antibody [EPR24184-52] (ab281563)

      HRG was immunoprecipitated from 0.35 mg of Human liver tissue lysate with ab281563 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed from the immunoprecipitate using ab281563 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366), was used as secondary antibody at 1/5000 dilution.


      Lane 1: Human liver tissue lysate 10µg (Input).
      Lane 2: ab281563 IP in Human liver tissue lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab281563 in Human liver tissue lysate.


      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 15 seconds.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)

      Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling HRG with ab281563 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on human tonsil. The section was incubated with ab281563 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

      Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HRG antibody [EPR24184-52] (ab281563)

      Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling HRG with ab281563 at 1/5000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control: no staining on human pancreas. The section was incubated with ab281563 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counter stained with Hematoxylin.

      Secondary antibody only control: Ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

      Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

    • Anti-HRG antibody [EPR24184-52] (ab281563)
      Anti-HRG antibody [EPR24184-52] (ab281563)

    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 (0)

    Publishing research using ab281563? Please let us know so that we can cite the reference in this datasheet.

    ab281563 has not yet been referenced specifically in any publications.

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