Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Fyn antibody [EPR19636] (ab184276)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19636] to Fyn
  • Suitable for: WB, IHC-P, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Fyn antibody [EPR19636]
    See all Fyn primary antibodies

  • Description

    Rabbit monoclonal [EPR19636] to Fyn

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Mouse Fyn aa 1-100. The exact sequence is proprietary.
    Database link: P39688

  • Positive control

    • WB: Human brain lysate; Ramos, HEK-293, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; P0 mouse brain lysate; P0 rat brain lysate; Rat spleen lysate. IHC-P: Human tonsil and cerebral cortex tissues; Mouse spleen tissue; Rat testis tissue. Flow Cyt: RAW 264.7 cells. IP: P0 mouse brain 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab184276 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 1/1000. Detects a band of approximately 61 kDa (predicted molecular weight: 61 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/50.
IP 1/40.

Target

  • Function

    Tyrosine-protein kinase implicated in the control of cell growth. Plays a role in the regulation of intracellular calcium levels, with isoform 2 showing the greater ability to mobilize cytoplasmic calcium in comparison to isoform 1. Required in brain development and mature brain function with important roles in the regulation of axon growth, axon guidance, and neurite extension. Blocks axon outgrowth and attraction induced by NTN1 by phosphorylating its receptor DDC. Phosphorylates RUNX3.

  • Tissue specificity

    Isoform 1 is highly expressed in the brain. Isoform 2 is expressed in cells of hemopoietic lineages, especially T lymphocytes.

  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. SRC subfamily.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
    Contains 1 SH3 domain.

  • Cellular localization

    Cell membrane. Present and active in lipid rafts. Present in cell body and along the process of mature and developing oligodendroyctes.
  • Information by UniProt

  • Database links

    see all

  • Form

    This protein is known to be similar in amino acid sequence to HCK (P08631), LCK (P06239), YES1 (P07947), SRC (P12931), and LYN (P07948). Therefore, cross-reactivity with these homologous proteins may be observed. We would be happy to provide immunogen alignment information upon request.

  • Alternative names

    • C syn protooncogene antibody
    • Fyn antibody
    • FYN oncogene related to SRC FGR YES antibody
    • FYN_HUMAN antibody
    • OKT3 induced calcium influx regulator antibody
    • P59 FYN antibody
    • p59-Fyn antibody
    • Protein tyrosine kinase fyn antibody
    • Proto oncogene tyrosine protein kinase fyn antibody
    • Proto-oncogene c-Fyn antibody
    • Proto-oncogene Syn antibody
    • Protooncogene Syn antibody
    • SLK antibody
    • Src like kinase antibody
    • Src yes related novel gene antibody
    • Src-like kinase antibody
    • Src/yes related novel antibody
    • SYN antibody
    • Tyrosine kinase p59fyn T antibody
    • Tyrosine kinase p59fyn(T) antibody
    • Tyrosine-protein kinase Fyn antibody
    see all

Images

  • Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T cell lysate
    Lane 2 : FYN knockout HEK-293T cell lysate
    Lane 3 : Human brain tissue lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 61 kDa
    Observed band size: 159 kDa
    why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab184276 observed at 60 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab184276 Anti-Fyn antibody [EPR19636] was shown to specifically react with Fyn in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266133 (knockout cell lysate ab257071) was used. Wild-type and Fyn knockout samples were subjected to SDS-PAGE. ab184276 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution

    Lane 1 : Human brain lysate
    Lane 2 : Ramos (human Burkitt's lymphoma cell line) whole cell lysate
    Lane 3 : HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate
    Lane 4 : P0 mouse brain lysate
    Lane 5 : P0 rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
    Lanes 2-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 61 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lanes 1 and 2: 3 minutes; Lanes 3,4 and 5: 8 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of human tonsil (PMID: 10523617). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)

    Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on human cerebral cortex (PMID: 7544314). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on T cells of mouse spleen (PMID: 7544314, PMID:10523617). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fyn antibody [EPR19636] (ab184276)

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling Fyn with ab184276 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on rat testis (PMID: 7544314). Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    Western blot - Anti-Fyn antibody [EPR19636] (ab184276)
    All lanes : Anti-Fyn antibody [EPR19636] (ab184276) at 1/1000 dilution

    Lane 1 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 3 : NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate
    Lane 4 : Rat spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 61 kDa
    Observed band size: 60 kDa why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Flow Cytometry - Anti-Fyn antibody [EPR19636] (ab184276)
    Flow Cytometry - Anti-Fyn antibody [EPR19636] (ab184276)

    Flow cytometric analysis of 4% paraformaldehyde-fixed RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) cell line labeling Fyn with ab184276 at 1/50 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-Fyn antibody [EPR19636] (ab184276)
    Immunoprecipitation - Anti-Fyn antibody [EPR19636] (ab184276)

    Fyn was immunoprecipitated from 0.35 mg of P0 mouse brain lysate with ab184276 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab184276 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: P0 mouse brain lysate 10 µg (Input). 

    Lane 2: ab184276 IP in P0 mouse brain lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab184276 in P0 mouse brain lysate.

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

    Exposure time: 3 minutes.

  • Anti-Fyn antibody [EPR19636] (ab184276)
    Anti-Fyn antibody [EPR19636] (ab184276)

References (1)

Publishing research using ab184276? Please let us know so that we can cite the reference in this datasheet.

ab184276 has been referenced in 1 publication.

  • Tang Y  et al. Bioinformatic analysis of differentially expressed genes and identification of key genes in EBV-transformed lymphoblasts. Biomed Pharmacother 116:108984 (2019). PubMed: 31129512

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