Overview

  • Product name

    Anti-Growth hormone receptor antibody
    See all Growth hormone receptor primary antibodies
  • Description

    Rabbit polyclonal to Growth hormone receptor
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Non human primates
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Growth hormone receptor.

    Read Abcam's proprietary immunogen policy (Peptide available as ab73395.)

  • Positive control

    • This antibody gave a positive result in the following whole cell lysates: A549, HeLa, DU 145

Properties

Applications

Our Abpromise guarantee covers the use of ab65304 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
ICC/IF Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 76 kDa (predicted molecular weight: 71 kDa).

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

  • 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

  • All lanes : Anti-Growth hormone receptor antibody (ab65304) at 1 µg/ml

    Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 71 kDa
    Observed band size: 76 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 56 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes


    Growth hormone receptor contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
  • ICC/IF image of ab65304 stained HepG2 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65304, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) HeLa, Hek293 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.

References

This product has been referenced in:

  • Beltran-Sastre V  et al. Tuneable endogenous mammalian target complementation via multiplexed plasmid-based recombineering. Sci Rep 5:17432 (2015). WB ; Human . Read more (PubMed: 26612112) »
See 1 Publication for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

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
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

Dr. Maïté Verreault

Verified customer

Submitted May 02 2013

Application
Western blot
Sample
Human Cell lysate - whole cell (Glioblastoma)
Loading amount
30 µg
Specification
Glioblastoma
Gel Running Conditions
Reduced Denaturing (4-15%)
Blocking step
Pierce TBS Super block Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 02 2013

Answer

Thank you for contacting us. The immunogen sequences for these antibodies are considered proprietary, but I can tell you that for both ab65304 and ab78426 the sequences are from human and do not overlap. Ab65304 was raised against an immunogen from within the range of amino acids 1-100, while ab78426 was raised against an immunogen from within the range of amino acids 101-150.
The immunogen for ab65304 shows very little homology with the sequence of the cow protein ab68383 and we would not expect it to detect ab68383 in WB. While the immunogen sequence for ab78426 is well conserved in the cow protein, we have no information about whether ab78426 will react with cow since it has not been tested.
We are still working to produce a new batch of 65304 that will pass our QC standards, but do not currently have an estimate as to when this antibody will be available. If you visit the online datasheet, there is an option to login to be notified when the product comes into stock. I hope this helps, please let me know if you need any additional information or assistance.

Read More

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