Recombinant
RabMAb

Recombinant Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)

Reviews (1)Q&A (4)References (3)

Overview

  • Product name

    Anti-Growth hormone receptor antibody [EPR5291(2)]
    See all Growth hormone receptor primary antibodies

  • Description

    Rabbit monoclonal [EPR5291(2)] to Growth hormone receptor

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Growth hormone receptor aa 550-650 (intracellular). The exact sequence is proprietary.

  • Positive control

    • WB: MCF-7, BxPC-3, and human fetal muscle, mouse brain and rat brain lysate. ICC/IF: HepG2 cells. Flow Cyt: MCF-7 cells.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab134078 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 1/2000 - 1/10000. Detects a band of approximately 140 kDa (predicted molecular weight: 72 kDa).
Flow Cyt 1/420.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
ICC/IF 1/200.
  • Application notes
    Is unsuitable for IHC-P.

    • Target

    • Function

      Receptor for pituitary gland growth hormone involved in regulating postnatal body growth. On ligand binding, couples to the JAK2/STAT5 pathway.
      The soluble form (GHBP) acts as a reservoir of growth hormone in plasma and may be a modulator/inhibitor of GH signaling.
      Isoform 2 up-regulates the production of GHBP and acts as a negative inhibitor of GH signaling.

    • Tissue specificity

      Expressed in various tissues with high expression in liver and skeletal muscle. Isoform 4 is predominantly expressed in kidney, bladder, adrenal gland and brain stem. Isoform 1 expression in placenta is predominant in chorion and decidua. Isoform 4 is highly expressed in placental villi. Isoform 2 is expressed in lung, stomach and muscle. Low levels in liver.

    • Involvement in disease

      Defects in GHR are a cause of Laron syndrome (LARS) [MIM:262500]. A severe form of growth hormone insensitivity characterized by growth impairment, short stature, dysfunctional growth hormone receptor, and failure to generate insulin-like growth factor I in response to growth hormone.
      Defects in GHR may be a cause of idiopathic short stature autosomal (ISSA) [MIM:604271]. Short stature is defined by a subnormal rate of growth.

    • Sequence similarities

      Belongs to the type I cytokine receptor family. Type 1 subfamily.
      Contains 1 fibronectin type-III domain.

    • Domain

      The WSXWS motif appears to be necessary for proper protein folding and thereby efficient intracellular transport and cell-surface receptor binding.
      The box 1 motif is required for JAK interaction and/or activation.
      The extracellular domain is the ligand-binding domain representing the growth hormone-binding protein (GHBP).
      The ubiquitination-dependent endocytosis motif (UbE) is required for recruitment of the ubiquitin conjugation system on to the receptor and for its internalization.

    • Post-translational
      modifications

      The soluble form (GHBP) is produced by phorbol ester-promoted proteolytic cleavage at the cell surface (shedding) by ADAM17/TACE. Shedding is inhibited by growth hormone (GH) binding to the receptor probably due to a conformational change in GHR rendering the receptor inaccessible to ADAM17.
      On GH binding, phosphorylated on tyrosine residues in the cytoplasmic domain by JAK2.
      On ligand binding, ubiquitinated on lysine residues in the cytoplasmic domain. This ubiquitination is not sufficient for GHR internalization.

    • Cellular localization

      Secreted; Cell membrane. On growth hormone binding, GHR is ubiquitinated, internalized, down-regulated and transported into a degradative or non-degradative pathway and Cell membrane. Remains fixed to the cell membrane and is not internalized.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • GH receptor antibody
      • GH-binding protein antibody
      • GHBP antibody
      • GHBP, included antibody
      • GHR antibody
      • GHR_HUMAN antibody
      • Growth hormone binding protein antibody
      • Growth hormone receptor antibody
      • Growth hormone receptor precursor antibody
      • Growth hormone-binding protein antibody
      • Growth hormone-binding protein, included antibody
      • Increased responsiveness to growth hormone, included antibody
      • Serum binding protein antibody
      • Serum-binding protein antibody
      • Somatotropin receptor antibody
      see all

    Images

    • Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078) at 1/2000 dilution (purified) + BxPC-3 cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 72 kDa
      Observed band size: 140 kDa
      why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078) at 1/10000 dilution (purified) + MCF-7 cell lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 72 kDa
      Observed band size: 140 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Western blot - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      All lanes : Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078) at 1/10000 dilution (purified)

      Lane 1 : Mouse brain
      Lane 2 : Rat brain

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 72 kDa
      Observed band size: 140 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Immunocytochemistry/ Immunofluorescence - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)

      Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Growth hormone receptor with purified ab134078 at 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

      Control: primary antibody (1/500) and secondary antibody ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/400).

    • Immunocytochemistry/ Immunofluorescence - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Immunocytochemistry/ Immunofluorescence - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)This image is courtesy of an anonymous Abreview

      ab134078 (unpurified) staining Growth hormone receptor in a human glioblastoma cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde,permeabilized with TBS Tween 0.5% and blocked for 60 minutes at 25°C. Samples were incubated with primary antibody (1/100) for 20 hour at 4°C. An Alexa Fluor® 594-conjugated Goat anti-rabbit IgG polyclonal(1/200) was used as the secondary antibody.

      See Abreview

    • Flow Cytometry - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Flow Cytometry - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)

      Flow cytometry analysis of MCF-7 cells labelling Growth hormone receptor with purified ab134078 at 1/420 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • Other - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Other - Anti-Growth hormone receptor antibody [EPR5291(2)] (ab134078)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    Publishing research using ab134078? Please let us know so that we can cite the reference in this datasheet.

    ab134078 has been referenced in 3 publications.

    • Basu R  et al. Targeting growth hormone receptor in human melanoma cells attenuates tumor progression and epithelial mesenchymal transition via suppression of multiple oncogenic pathways. Oncotarget 8:21579-21598 (2017). WB, ICC/IF ; Human . PubMed: 28223541
    • Lev A  et al. Preclinical rationale for combination of crizotinib with mitomycin C for the treatment of advanced colorectal cancer. Cancer Biol Ther 18:694-704 (2017). PubMed: 28886275
    • Lee EH  et al. Immunogenomics reveal molecular circuits of diclofenac induced liver injury in mice. Oncotarget 7:14983-5017 (2016). PubMed: 26934552

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Question

    I would like to know the difference between this product (ab134078) and a different polyclonal GHR antibody (ab65304). Specifically, why the band observed using ab65304 for western blot is 76kDa (much closer to the predicted size of 71 kDa), while this monoclonal antibody detects a band around 150kDa. Are both of these antibodies validated to detect only GHR? Please let me know any information you may have about this.

    Read More
    Answer

    Growth hormone receptor is about 72 kDa before any post-translation modifications. (See the section titled Sequences, halfway down the page of the UniProt record for human GHR at this link: http://www.uniprot.org/uniprot/P10912 ).

    But subsequent glycosylation causes the protein to migrate in SDS-PAGE gels at higher molecular weights, according to several papers. For instance, the paper at this link, http://joe.endocrinology-journals.org/content/198/1/71.full.pdf , has this note on page78.

    Based on its amino acid sequence, the nascent hGHR is
    predicted to be ˜70 kDa in size while, when resolved on
    SDS-PAGE gels under reducing conditions, the mature
    hGHR migrates at 100–140 kDa (Hocquette et al. 1989,
    1990, Alele et al. 1998, Zhang et al. 2001, Cowan et al. 2005).
    These differences are due primarily to the presence of five
    asparagine-linked glycosylation sites within the extracellular
    domain, a variable number of which are glycosylated in the
    mature hGHR (Harding et al. 1994).

    J Endocrinol. 2008 Jul;198(1):71-82. PMID: 18420710

    Neither of the two antibodies, ab65304 and ab134078, have been tested for reactivity with purified receptor, or with knockout/knockdown samples to demonstrate specificity for GHR. At least one customer reported detection of two bands at 110 kDa and 71 kDa in a blot of human glioblasoma lysate stained with ab65304. The review is at this link, https://www.abcam.com/Growth-hormone-receptor-antibody-ab65304/reviews/35503 .

    There is a possibility both antibodies detect the receptor but they were unfortunately not tested against the same samples, and we do not know if the different results are a consequence of different amounts of glycosylation among those samples or, more likely, different affinities of the antibodies for glycosylated receptor. I would choose ab134078 over ab65304, given that the western result agrees with the observations mentioned in the references above. Our guarantee, described at https://www.abcam.com/content/abpromise-guarantee, applies to all the species and applications listed on the datasheet.

    Read More

    Question

    What is least range of  homology if one antibody have cross activity with other organisms ? Is 73% not enough?Best Wishes!

    Read More
    Answer

    While an antibody created from an immunogen with 73% homology with a target of interest may have reactivity, Abcam does not recommend use of a product which has less than 85% homology.

    Read More

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Growth hormone receptor antibody [EPR5291(2)]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Glioblastoma)
    Specification
    Glioblastoma
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - TBS Tween 0.5%
    Blocking step
    (agent) for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Feb 13 2013

    Question

    Inquiry: Bonjour, Nous avons bien reçu l'anticorps. Cependant je remarque qu'en western blot, il est indiqué sur votre fiche technique que la bande sort à 140kDa. J'obtenais toutefois une bande de façon consistante à 70kDa avec l'autre anticorps que nous utilisions. Pourquoi est-ce qu'il aurait une telle différence entre les deux anticorps s'ils sont supposés reconnaître la même protéine?

    Read More
    Answer

    Merci de nous avoir contactés.

    Le laboratoire Epitomics, source du monoclonal de lapin ab134078, confirme la spécificité de cet anticorps en s'appuyant sur les publications suivantes où la forme glycosylée de la GHR est décrite pour migrer à 120 ou 140 kDa :

    http://joe.endocrinology-journals.org/content/198/1/71.full.pdf
    http://joe.endocrinology-journals.org/cgi/content-nw/full/195/1/49/F3
    http://joe.endocrinology-journals.org/content/181/2/281.full.pdf
    http://datasheets.scbt.com/sc-69880.pdf
    http://mend.endojournals.org/content/17/11/2211.full

    Ceci est en effet en contradiction avec les blots obtenus avec les autres anti-GHR de notre catalogue. Je vais donc demander à mes collègues en charge de ces anticorps de bien vouloir vérifier et si possible expliquer ces différences.

    Je vous contacterai de nouveau quand j'aurai reçu une réponse de mes collaborateurs.
    Merci pour votre patience.

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    Question

    Nous choisirons le produit ab134078 (https://www.abcam.com/ab134078) en remplacement du produit non disponible et nous le testerons en immunofluorescence.

    Read More
    Answer

    Code de remise : xxxxxxxxx
    Date d’expiration : 30/11/12

    Je suis ravie que vous souhaitiez participer a notre offre et tester ab134078 en immunohistochimie. Nous avons passé votre commande et le code de réduction correspondant est xxxxxxxxxx. Ce code de réduction est utilisable sous 120 jours et pour l’achat d’un autre anticorps primaire.

    Vous devez dans un premier temps soumettre une Abreview avec vos résultats en immunohistochimie en notant ce code dans la section « Additional notes ». Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de ab134078.

    Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.

    Termes et Conditions : www.abcam.com/collaborationdiscount.

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