Recombinant 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: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-JAK3 antibody [EP909Y]
See all JAK3 primary antibodies -
Description
Rabbit monoclonal [EP909Y] to JAK3 -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
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 (intra): 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
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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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...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EP909Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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.
Application | Abreviews | Notes |
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Flow Cyt (Intra) |
1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/2000. Predicted molecular weight: 125 kDa.
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IP |
1/60.
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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.
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ICC/IF | (1) |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/2000. Predicted molecular weight: 125 kDa. |
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
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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
modificationsTyrosine phosphorylated in response to IL-2 and IL-4. -
Cellular localization
Endomembrane system. Wholly intracellular, possibly membrane associated. - Information by UniProt
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Database links
- Entrez Gene: 3718 Human
- Omim: 600173 Human
- SwissProt: P52333 Human
- Unigene: 515247 Human
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Alternative names
- EC 2.7.10.2 antibody
- JAK 3 antibody
- JAK L antibody
see all
Images
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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 minutesBlocking/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 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.
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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 secondsBlocking and diluting buffer: 5% NFDM/TBST.
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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) 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).
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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).
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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 -
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 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 minutes at 22°C. Isotype control antibody (black line) was rabbit IgG (1µg/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.
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.
Datasheets and documents
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SDS download
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Datasheet download
References (10)
ab45141 has been referenced in 10 publications.
- Xu L et al. FABP4 activates the JAK2/STAT2 pathway via Rap1a in the homocysteine-induced macrophage inflammatory response in ApoE-/- mice atherosclerosis. Lab Invest 102:25-37 (2022). PubMed: 34725437
- Li C et al. Suppressive function of bone marrow-derived mesenchymal stem cell-derived exosomal microRNA-187 in prostate cancer. Cancer Biol Ther 23:1-14 (2022). PubMed: 36245088
- Zhang J et al. CD105 expression is associated with invasive capacity in ovarian cancer and promotes invasiveness by inhibiting NDRG1 and regulating the epithelial-mesenchymal transition. Am J Transl Res 13:12461-12479 (2021). PubMed: 34956466
- Vadivel CK et al. JAK3 Is Expressed in the Nucleus of Malignant T Cells in Cutaneous T Cell Lymphoma (CTCL). Cancers (Basel) 13:N/A (2021). PubMed: 33466582
- Jiang H et al. Transcriptome-Wide High-Throughput m6A Sequencing of Differential m6A Methylation Patterns in the Human Rheumatoid Arthritis Fibroblast-Like Synoviocytes Cell Line MH7A. J Inflamm Res 14:575-586 (2021). PubMed: 33658830
- Liang Y et al. Curcumin inhibits the viability, migration and invasion of papillary thyroid cancer cells by regulating the miR-301a-3p/STAT3 axis. Exp Ther Med 22:875 (2021). PubMed: 34194553
- Yang J & Xie X Tofacitinib protects intestinal epithelial cells against oxygen-glucose deprivation/reoxygenation injury by inhibiting the JAK/STAT3 signaling pathway. Exp Ther Med 22:1108 (2021). PubMed: 34504562
- Chen S et al. N-myc Downstream-Regulated Gene 2 (NDRG2) Promotes Bone Morphogenetic Protein 2 (BMP2)-Induced Osteoblastic Differentiation and Calcification by Janus Kinase 3 (JAK3)/Signal Transducer and Activator of Transcription 3 (STAT3) Signaling Pathway. Med Sci Monit 26:e918541 (2020). PubMed: 31911574
- Liu B & Ren B MiR-1193 represses the proliferation and induces the apoptosis of interleukin-1ß-treated fibroblast-like synoviocytes via targeting JAK3. Int J Rheum Dis 23:1066-1075 (2020). PubMed: 32597556
- 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