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Signal Transduction Protein Phosphorylation Tyrosine Kinases Other
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RecombinantRabMAb

Recombinant Anti-JAK3 antibody [EP909Y] (ab45141)

  • Datasheet
  • SDS
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Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Immunocytochemistry/ Immunofluorescence - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Immunoprecipitation - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Flow Cytometry - Anti-JAK3 antibody [EP909Y] (ab45141)
  • Anti-JAK3 antibody [EP909Y] (ab45141)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP909Y] to JAK3
  • Suitable for: WB, Flow Cyt, IP, IHC-P, ICC/IF
  • Reacts with: Human

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Overview

  • Product name

    Anti-JAK3 antibody [EP909Y]
    See all JAK3 primary antibodies
  • Description

    Rabbit monoclonal [EP909Y] to JAK3
  • Host species

    Rabbit
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human JAK3 aa 1100 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P52333

  • Positive control

    • Flow Cyt: Jurkat cells. WB: TF-1 cell lysate, HEL, KARPAS-299, HaCaT. IHC-P: Human NK cell lymphoma, Human large B cell lymphoma ICC/IF: KARPAS-299IP: TF-1 whole cell lysate
  • General notes

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

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Tissue culture supernatant
  • Clonality

    Monoclonal
  • Clone number

    EP909Y
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Protein Phosphorylation
    • Tyrosine Kinases
    • Other
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • STATs
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • STATs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Tyrosine kinases
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Signal transducers

Associated products

  • Alternative Versions

    • Anti-JAK3 antibody [EP909Y] - BSA and Azide free (ab232005)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human JAK3 protein (ab89982)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab45141 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Human
WB
Human
Application Abreviews Notes
WB
1/2000. Predicted molecular weight: 125 kDa.
Flow Cyt
1/100.

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

IP
1/60.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF (1)
Use at an assay dependent concentration.
Notes
WB
1/2000. Predicted molecular weight: 125 kDa.
Flow Cyt
1/100.

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

IP
1/60.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.

Target

  • Function

    Tyrosine kinase of the non-receptor type, involved in the interleukin-2 and interleukin-4 signaling pathway. Phosphorylates STAT6, IRS1, IRS2 and PI3K.
  • Tissue specificity

    In NK cells and an NK-like cell line but not in resting T-cells or in other tissues. The S-form is more commonly seen in hematopoietic lines, whereas the B-form is detected in cells both of hematopoietic and epithelial origins.
  • Involvement in disease

    Defects in JAK3 are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-negative (T(-)B(+)NK(-) SCID) [MIM:600802]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
  • Sequence similarities

    Belongs to the protein kinase superfamily. Tyr protein kinase family. JAK subfamily.
    Contains 1 FERM domain.
    Contains 1 protein kinase domain.
    Contains 1 SH2 domain.
  • Domain

    Possesses two phosphotransferase domains. The second one probably contains the catalytic domain (By similarity), while the presence of slight differences suggest a different role for domain 1.
  • Post-translational
    modifications

    Tyrosine phosphorylated in response to IL-2 and IL-4.
  • Cellular localization

    Endomembrane system. Wholly intracellular, possibly membrane associated.
  • Target information above from: UniProt accession P52333 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 3718 Human
    • Omim: 600173 Human
    • SwissProt: P52333 Human
    • Unigene: 515247 Human
    • Alternative names

      • EC 2.7.10.2 antibody
      • JAK 3 antibody
      • JAK L antibody
      • JAK-3 antibody
      • Jak3 antibody
      • JAK3 HUMAN antibody
      • JAK3_HUMAN antibody
      • JAKL antibody
      • Janus kinase 3 (a protein tyrosine kinase, leukocyte) antibody
      • Janus kinase 3 antibody
      • Janus Kinase3 antibody
      • L JAK antibody
      • L-JAK antibody
      • Leukocyte janus kinase antibody
      • LJAK antibody
      • Protein tyrosine kinase leukocyte antibody
      • Tyrosine protein kinase JAK3 antibody
      • Tyrosine-protein kinase JAK3 antibody
      see all

    Images

    • Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      All lanes : Anti-JAK3 antibody [EP909Y] (ab45141) at 1/1000 dilution

      Lane 1 : TF-1 (Human Erythroleukemia erythroblast) whole cell lysate
      Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 125 kDa
      Observed band size: 125 kDa


      Exposure time: 3 minutes


      Blocking/Diluting buffer and concentration 5% NFDM/TBST

      The expression profile observed in HaCaT is consistent with the literature (PMID: 11709700).
      Negative control: HaCaT (PMID: 11709700)

    • Immunocytochemistry/ Immunofluorescence - Anti-JAK3 antibody [EP909Y] (ab45141)
      Immunocytochemistry/ Immunofluorescence - Anti-JAK3 antibody [EP909Y] (ab45141)

      Immunocytochemistry/Immunofluorescence analysis of KARPAS-299 (human anaplastic large cell lymphoma) labelling JAK3 with ab45141 at a dilution of 1:100 dilution (12 µg/ml). Cells were fixed with 100% Methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) (1:1000 dilution (2 µg/ml)) was used as the secondary antibody. The cells were co-stained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Nuclei counterstained with DAPI (blue). Control: PBS instead of the primary antibody.

    • Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      All lanes : Anti-JAK3 antibody [EP909Y] (ab45141) at 1.21 µg/ml

      Lane 1 : KARPAS-299 (Human anaplastic large cell lymphoma ) whole cell lysate
      Lane 2 : HEL (Human Erythroleukemia erythroblast ) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 125 kDa
      Observed band size: 125 kDa


      Exposure time: 26 seconds


      Blocking and diluting buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-JAK3 antibody [EP909Y] (ab45141)
      Immunoprecipitation - Anti-JAK3 antibody [EP909Y] (ab45141)

      Lane 1 (input): TF-1 (Human Erythroleukemia erythroblast) whole cell lysate, 10µg
      Lane 2 (+): TF-1 whole cell lysate
      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab45141 in TF-1 whole cell lysate.

      ab45141 at 1/60 immunoprecipitating JAK3 in TF-1 whole cell lysate. For western blotting, ab45141 was used as a primary antibody at 1/500 dilution (2.42 µg/ml). ab131366, VeriBlot for IP Detection Reagent was used for detection at 1/1000 dilution. Blocking and diluting buffer used was 5% NFDM/TBST.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)

      Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of Human NK cell lymphoma tissue sections labeling JAK3 using purified ab45141. Samples were incubated the primary antibody at 1:2000 dilution (0.60 μg/ml). Hematoxylin was used as a counterstain. PBS instead of primary antibody was used for negative control. A ready to use ImmunoHistoProbe one step HRP Polymer at 1:0 dilution was used as the secondary antibody. Heatm mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JAK3 antibody [EP909Y] (ab45141)

      Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of Human large B cell lymphoma tissue sections labeling JAK3 using purified ab45141. Samples were incubated the primary antibody at 1:2000 dilution (0.60 μg/ml). Hematoxylin was used as a counterstain. PBS instead of primary anitbody was used for negative control. A ready to use ImmunoHistoProbe one step HRP Polymer at 1:0 dilution was used as the secondary antibody. Heatm mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      Western blot - Anti-JAK3 antibody [EP909Y] (ab45141)
      Anti-JAK3 antibody [EP909Y] (ab45141) at 1/2000 dilution + TF-1 (Human bone marrow erythroleukemia cell line) cell lysate

      Predicted band size: 125 kDa
      Observed band size: 125 kDa

    • Flow Cytometry - Anti-JAK3 antibody [EP909Y] (ab45141)
      Flow Cytometry - Anti-JAK3 antibody [EP909Y] (ab45141)

      Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab45141 (red line). The cells were fixed with 80% methanol (5 minutes) and then permeabilized with 0.1% PBS-Tween for 20 minutes. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab45141, 1/100 dilution) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was rabbit IgG (1 µg/1x106 cells ) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 4% paraformaldehyde (10 minutes)/permeabilized in 0.1% PBS-Tween used under the same conditions.

    • Anti-JAK3 antibody [EP909Y] (ab45141)
      Anti-JAK3 antibody [EP909Y] (ab45141)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (1)

    Publishing research using ab45141? Please let us know so that we can cite the reference in this datasheet.

    ab45141 has been referenced in 1 publication.

    • Wefers C  et al. Different Lipid Regulation in Ovarian Cancer: Inhibition of the Immune System. Int J Mol Sci 19:N/A (2018). PubMed: 29342108

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    Immunocytochemistry/ Immunofluorescence abreview for Anti-JAK3 antibody [EP909Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (T-cell line from skin)
    Permeabilization
    Yes - Ice cold Methanol
    Specification
    T-cell line from skin
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Aug 09 2019

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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