FGFR3 Recombinant Rabbit Monoclonal Antibody [JM81-10] - BSA and Azide free
Catalog# HA750379
FGFR3 Recombinant Rabbit Monoclonal Antibody [JM81-10] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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FC
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IP
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Human
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unconjugated
Overview
Product Name
FGFR3 Recombinant Rabbit Monoclonal Antibody [JM81-10] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within Human FGFR3 aa 719-806 / 806.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Target Molecular Weight
Predicted band size: 88 kDa
Positive Control
HepG2 cell lysate, HEK-293 cell lysate, LNCaP cell lysate, A549 cell lysate, Hela, MCF-7, SH-SY5Y, human breast carcinoma tissue, human kidney tissue, HepG2.
Conjugation
unconjugated
Clone Number
JM81-10
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
Function
Acidic and basic fibroblast growth factors (FGFs) are members of a family of multifunctional polypeptide growth factors that stimulate proliferation of cells of mesenchymal, epithelial and neuroectodermal origin. Like other growth factors, FGFs act by binding and activating specific cell surface receptors. These include the Flg receptor or FGFR-1, the Bek receptor or FGFR-2, FGFR-3, FGFR-4, FGFR-5 and FGFR-6. These receptors usually contain an extracellular ligand-binding region containing three immunoglobulin-like domains, a transmembrane domain and a cytoplasmic tyrosine kinase domain. The gene encoding human FGFR-3 maps to chromosome 4p16 and is alternatively spliced to produce three isoforms that are expressed in brain, kidney and testis. Defects in FGFR-3 are associated with several diseases, including Crouzon syndrome, achondroplasia, thanatophoric dysplasia, craniosynostosis adelaide type and hypochondroplasia. Mutations in FGFR-3 are also a cause of some bladder and cervical cancers.
Background References
1. Wolk K et al. Deficient cutaneous antibacterial competence in cutaneous T-cell lymphomas: role of Th2-mediated biased Th17 function. Clin Cancer Res 20:5507-16 (2014).
2. Chung TW et al. Lipocalin-2 elicited by advanced glycation end-products promotes the migration of vascular smooth muscle cells. Biochim Biophys Acta 1833:3386-95 (2013).
Sequence Similarity
Belongs to the protein kinase superfamily. Tyr protein kinase family. Fibroblast growth factor receptor subfamily.
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.
Post-translational Modification
Autophosphorylated. Binding of FGF family members together with heparan sulfate proteoglycan or heparin promotes receptor dimerization and autophosphorylation on tyrosine residues. Autophosphorylation occurs in trans between the two FGFR molecules present in the dimer. Phosphorylation at Tyr-724 is essential for stimulation of cell proliferation and activation of PIK3R1, STAT1 and MAP kinase signaling. Phosphorylation at Tyr-760 is required for interaction with PIK3R1 and PLCG1.; Ubiquitinated. Is rapidly ubiquitinated after ligand binding and autophosphorylation, leading to receptor internalization and degradation. Subject to both proteasomal and lysosomal degradation.; N-glycosylated in the endoplasmic reticulum. The N-glycan chains undergo further maturation to an Endo H-resistant form in the Golgi apparatus.
Subcellular Location
Endoplasmic reticulum, Cell membrane, Cytoplasmic vesicle, Secreted.
UNIPROT
Synonyms
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
ExpandImages
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This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Western blot analysis of FGFR3 on different lysates with Rabbit anti-FGFR3 antibody (ET1703-36) at 1/5,000 dilution.
Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: HEK-293 cell lysate (15 µg/Lane)
Lane 3: LNCaP cell lysate (15 µg/Lane)
Lane 4: A549 cell lysate (15 µg/Lane)
Predicted band size: 88 kDa
Observed band size: 130 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-36) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Western blot analysis of FGFR3 on different lysates with Rabbit anti-FGFR3 antibody (ET1703-36) at 1/1,000 dilution.
Lane 1: HEK293-si NT cell lysate
Lane 2: HEK293-si FGFR3 cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 88 kDa
Observed band size: 130 kDa
Exposure time: 3 minutes;
4-20% SDS-PAGE gel.
ET1703-36 was shown to specifically react with FGFR3 in HEK293-si NT cells. Weakened band was observed when HEK293-si FGFR3 sample was tested. HEK293-si NT and HEK293-si FGFR3 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1703-36, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature. -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of HepG2 cells labeling FGFR3 with Rabbit anti-FGFR3 antibody (ET1703-36) at 1/100 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-FGFR3 antibody (ET1703-36) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
ICC staining of FGFR3 in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-36, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
ICC staining of FGFR3 in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-36, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
ICC staining of FGFR3 in SH-SY5Y cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-36, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-FGFR3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-36, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-FGFR3 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-36, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of FGFR3 was done on HepG2 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-36, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
This data was developed using ET1703-36, the same antibody clone in a different buffer formulation.
FGFR3 was immunoprecipitated in 0.2mg HepG2 cell lysate with ET1703-36 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1703-36 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HepG2 cell lysate (input)
Lane 2: ET1703-36 IP in HepG2 cell lysate
Lane 3: Rabbit IgG instead of ET1703-36 in HepG2 cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 4 mintues
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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