Recombinant
RabMAb

Anti-FGFR3 (phospho Y724) antibody [EPR2281(3)] (ab155960)

Overview

  • Product name
    Anti-FGFR3 (phospho Y724) antibody [EPR2281(3)]
    See all FGFR3 primary antibodies
  • Description
    Rabbit monoclonal [EPR2281(3)] to FGFR3 (phospho Y724)
  • Host species
    Rabbit
  • Specificity
    ab155960 only detects FGFR3 when phosphorylated at tyrosine 724.
  • Tested applications
    Suitable for: Flow Cyt, WB, ICC/IF, IPmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human FGFR3 (phospho Y724).
    Database link: P22607

  • Positive control
    • WB: MCF-7 lysate treated with pervanadate. ICC/IF: MCF-7 cells treated with pervanadate. IP: MCF-7 cells treated with pervanadate. FC: MCF-7 cells
  • General notes

    Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

     

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab155960 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
Flow Cyt 1/50.
WB 1/1000 - 1/10000. Predicted molecular weight: 88 kDa.

 

ICC/IF 1/80 - 1/300.

 

IP 1/15 - 1/50.

 

  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Receptor for acidic and basic fibroblast growth factors. Preferentially binds FGF1.
    • Tissue specificity
      Expressed in brain, kidney and testis. Very low or no expression in spleen, heart, and muscle. In 20- to 22-week old fetuses it is expressed at high level in kidney, lung, small intestine and brain, and to a lower degree in spleen, liver, and muscle. Isoform 2 is detected in epithelial cells. Isoform 1 is not detected in epithelial cells. Isoform 1 and isoform 2 are detected in fibroblastic cells.
    • Involvement in disease
      Defects in FGFR3 are the cause of achondroplasia (ACH) [MIM:100800]. ACH is an autosomal dominant disease and is the most frequent form of short-limb dwarfism. It is characterized by a long, narrow trunk, short extremities, particularly in the proximal (rhizomelic) segments, a large head with frontal bossing, hypoplasia of the midface and a trident configuration of the hands.
      Defects in FGFR3 are the cause of Crouzon syndrome with acanthosis nigricans (CAN) [MIM:612247]. Classic Crouzon disease which is caused by mutations in the FGFR2 gene is characterized by craniosynostosis (premature fusion of the skull sutures), and facial hypoplasia. Crouzon syndrome with acanthosis nigricans (a skin disorder characterized by pigmentation anomalies), CAN, is considered to be an independent disorder from classic Crouzon syndrome. CAN is characterized by additional more severe physical manifestation, such as Chiari malformation, hydrocephalus, and atresia or stenosis of the choanas, and is caused by a specific mutation (Ala-391 to Glu) in the transmembrane domain of FGFR3. It is proposed to have an autosomal dominant mode of inheritance.
      Defects in FGFR3 are a cause of thanatophoric dysplasia type (TD) [MIM:187600, 187601]; also known as thanatophoric dwarfism or platyspondylic lethal skeletal dysplasia Sand Diego type (PLSD-SD). TD is the most common neonatal lethal skeletal dysplasia. Affected individuals display features similar to those seen in homozygous achondroplasia. It causes severe shortening of the limbs with macrocephaly, narrow thorax and short ribs. In the most common subtype, TD1, femur are curved, while in TD2, straight femurs are associated with cloverleaf skull. Mutations affecting different functional domains of FGFR3 cause different forms of this lethal disorder.
      Defects in FGFR3 are a cause of hypochondroplasia (HCH) [MIM:146000]. HCH is an autosomal dominant disease and is characterized by disproportionate short stature. It resembles achondroplasia, but with a less severe phenotype.
      Defects in FGFR3 are a cause of susceptibility to bladder cancer (BLC) [MIM:109800]. A malignancy originating in tissues of the urinary bladder. It often presents with multiple tumors appearing at different times and at different sites in the bladder. Most bladder cancers are transitional cell carcinomas. They begin in cells that normally make up the inner lining of the bladder. Other types of bladder cancer include squamous cell carcinoma (cancer that begins in thin, flat cells) and adenocarcinoma (cancer that begins in cells that make and release mucus and other fluids). Bladder cancer is a complex disorder with both genetic and environmental influences. Note=Somatic mutations can constitutively activate FGFR3.
      Defects in FGFR3 are a cause of cervical cancer (CERCA) [MIM:603956]. A malignant neoplasm of the cervix, typically originating from a dysplastic or premalignant lesion previously present at the active squamocolumnar junction. The transformation from mild dysplastic to invasive carcinoma generally occurs slowly within several years, although the rate of this process varies widely. Carcinoma in situ is particularly known to precede invasive cervical cancer in most cases. Cervical cancer is strongly associated with infection by oncogenic types of human papillomavirus.
      Defects in FGFR3 are the cause of camptodactyly tall stature and hearing loss syndrome (CATSHL syndrome) [MIM:610474]. CATSHL syndrome is an autosomal dominant syndrome characterized by permanent and irreducible flexion of one or more fingers of the hand and/or feet, tall stature, scoliosis and/or a pectus excavatum, and hearing loss. Affected individuals have developmental delay and/or mental retardation, and several of these have microcephaly. Radiographic findings included tall vertebral bodies with irregular borders and broad femoral metaphyses with long tubular shafts. On audiological exam, each tested member have bilateral sensorineural hearing loss and absent otoacoustic emissions. The hearing loss was congenital or developed in early infancy, progressed variably in early childhood, and range from mild to severe. Computed tomography and magnetic resonance imaging reveal that the brain, middle ear, and inner ear are structurally normal.
      Defects in FGFR3 are a cause of multiple myeloma (MM) [MIM:254500]. MM is a malignant tumor of plasma cells usually arising in the bone marrow and characterized by diffuse involvement of the skeletal system, hyperglobulinemia, Bence-Jones proteinuria and anemia. Complications of multiple myeloma are bone pain, hypercalcemia, renal failure and spinal cord compression. The aberrant antibodies that are produced lead to impaired humoral immunity and patients have a high prevalence of infection. Amyloidosis may develop in some patients. Multiple myeloma is part of a spectrum of diseases ranging from monoclonal gammopathy of unknown significance (MGUS) to plasma cell leukemia. Note=A chromosomal aberration involving FGFR3 is found in multiple myeloma. Translocation t(4;14)(p16.3;q32.3) with the IgH locus.
      Defects in FGFR3 are a cause of lacrimo-auriculo-dento-digital syndrome (LADDS) [MIM:149730]; also known as Levy-Hollister syndrome. LADDS is a form of ectodermal dysplasia, a heterogeneous group of disorders due to abnormal development of two or more ectodermal structures. LADDS is an autosomal dominant syndrome characterized by aplastic/hypoplastic lacrimal and salivary glands and ducts, cup-shaped ears, hearing loss, hypodontia and enamel hypoplasia, and distal limb segments anomalies. In addition to these cardinal features, facial dysmorphism, malformations of the kidney and respiratory system and abnormal genitalia have been reported. Craniosynostosis and severe syndactyly are not observed.
      Defects in FGFR3 are a cause of keratinocytic non-epidermolytic nevus (KNEN) [MIM:162900]; also known as pigmented moles. Epidermal nevi of the common, non-organoid and non-epidermolytic type are benign skin lesions and may vary in their extent from a single (usually linear) lesion to widespread and systematized involvement. They may be present at birth or develop early during childhood.
      Defects in FGFR3 are a cause of Muenke syndrome (MNKS) [MIM:602849]; also known as Muenke non-syndromic coronal craniosynostosis. MNKS is a condition characterized by premature closure of coronal suture of skull during development (coronal craniosynostosis), which affects the shape of the head and face. It may be uni- or bilateral. When bilateral, it is characterized by a skull with a small antero-posterior diameter (brachycephaly), often with a decrease in the depth of the orbits and hypoplasia of the maxillae. Unilateral closure of the coronal sutures leads to flattening of the orbit on the involved side (plagiocephaly). The intellect is normal. In addition to coronal craniosynostosis some affected individuals show skeletal abnormalities of hands and feet, sensorineural hearing loss, mental retardation and respiratory insufficiency.
      Defects in FGFR3 are a cause of keratosis seborrheic (KERSEB) [MIM:182000]. A common benign skin tumor. Seborrheic keratoses usually begin with the appearance of one or more sharply defined, light brown, flat macules. The lesions may be sparse or numerous. As they initially grow, they develop a velvety to finely verrucous surface, followed by an uneven warty surface with multiple plugged follicles and a dull or lackluster appearance.
    • Sequence similarities
      Belongs to the protein kinase superfamily. Tyr protein kinase family. Fibroblast growth factor receptor subfamily.
      Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
      Contains 1 protein kinase domain.
    • Cellular localization
      Membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • ACH antibody
      • CD 333 antibody
      • CD333 antibody
      • CD333 antigen antibody
      • CEK 2 antibody
      • CEK2 antibody
      • FGFR 3 antibody
      • FGFR-3 antibody
      • FGFR3 antibody
      • FGFR3_HUMAN antibody
      • Fibroblast growth factor receptor 3 (achondroplasia thanatophoric dwarfism) antibody
      • Fibroblast growth factor receptor 3 antibody
      • Heparin binding growth factor receptor antibody
      • HSFGFR3EX antibody
      • Hydroxyaryl protein kinase antibody
      • JTK 4 antibody
      • JTK4 antibody
      • MFR 3 antibody
      • SAM 3 antibody
      • Tyrosine kinase JTK 4 antibody
      • Tyrosine kinase JTK4 antibody
      • Z FGFR 3 antibody
      see all

    Images

    • All lanes : Anti-FGFR3 (phospho Y724) antibody [EPR2281(3)] (ab155960) at 1/20000 dilution (purified)

      Lane 1 : MCF-7 cell lysate - untreated
      Lane 2 : MCF-7 cell lysate - treated with pervanadate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 88 kDa
      Observed band size: 96 kDa (why is the actual band size different from the predicted?)



      Blocking and dilution buffer: 5% NFDM/TBST.

    • Flow Cytometry analysis of MCF-7 (human breast carcinoma) treated with 1 mM pervanadate for 30 minutes cells labeling FGFR3 (phospho Y724) with purified ab155960 at 1/50 dilution (red). The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. Green shows untreated MCF-7 (human breast carcinoma) cells. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

    • Immunocytochemistry/Immunofluorescence analysis of untreated, Per treated and Per + LP treated MCF-7 cells labelling FGFR3 (phospho Y724) with ab155960 (left) and FGFR3 with ab137084 (right) both at a dilution of 1/200.

      Cells were fixed with 100% methanol. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      The image shows increased cytoplasmic staining after Pervanadate (1 mM, 30 min) treatment on MCF7 cells. The LP treatment decreased the cytoplasmic staining caused by Pervanadate.

      ab137084 was used as a Pan control for ab155960. The results showed cytoplasmic staining on untreated, Per treated and Per + LP treated MCF7 cells.

    • All lanes : Anti-FGFR3 (phospho Y724) antibody [EPR2281(3)] (ab155960) at 1/10000 dilution (unpurified)

      Lane 1 : MCF-7 whole cell lysate - treated with pervanadate
      Lane 2 : MCF-7 whole cell lysate - untreated

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 88 kDa
      Observed band size: 98 kDa (why is the actual band size different from the predicted?)


      Exposure time: 30 seconds


      Blocking and dilution buffer: 5% NFDM/TBST.

    • Dot blot analysis of FGFR3 (pY724) phospho peptide (lane 1) and FGFR3 non-phospho peptide (lane 2) labelling FGFR2 (phospho Y724) with ab155960 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 10 seconds.

    • ab155960 (unpurified) at 1/15 immunoprecipitating FGFR3 (phospho Y724) in MCF-7 cells treated with pervanadate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • ab155960 (purified) at 1/50 immunoprecipitating FGFR3 (phospho Y724) in MCF-7 cells treated with pervanadate. For western blotting, a peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/1000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    References

    This product has been referenced in:
    • Datta J  et al. Akt Activation Mediates Acquired Resistance to Fibroblast Growth Factor Receptor Inhibitor BGJ398. Mol Cancer Ther 16:614-624 (2017). Read more (PubMed: 28255027) »
    • Takata N  et al. Self-patterning of rostral-caudal neuroectoderm requires dual role of Fgf signaling for localized Wnt antagonism. Nat Commun 8:1339 (2017). Read more (PubMed: 29109536) »

    See all 4 Publications for this product

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (lung)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    25 µg
    Treatment
    FGF19 (20ng/ml) for 60 minutes
    Specification
    lung
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted May 25 2017



    I am happy to confirm that serum starved (overnight) MCF-7 cells were incubated with 1mM pervanadate for 20 minutes at 37℃ to generated the sample we show in the WB image on the datasheet of ab155960.

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

    Sign up