Recombinant Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18841-95] to JNK1+JNK2+JNK3
- Suitable for: WB, ICC/IF, Flow Cyt, IP
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95]
See all JNK1+JNK2+JNK3 primary antibodies -
Description
Rabbit monoclonal [EPR18841-95] to JNK1+JNK2+JNK3 -
Host species
Rabbit -
Tested applications
Suitable for: WB, ICC/IF, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Human JNK1+JNK2+JNK3 aa 150-400. The exact sequence is proprietary. SwissProt ID: P45984 = JNK2 and P53779 = JNK3.
Database link: P45983 -
Positive control
- WB: Human JNK1, JNK2 and JNK3 full length recombinant proteins; Human fetal liver, fetal heart and fetal kidney lysates; Jurkat, HeLa, K562, MCF7, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Rat brain, kidney and spleen lysates. ICC/IF: HeLa and NIH/3T3 cell lines. Flow Cyt: Jurkat and HeLa cell lines. IP: HeLa 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.
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
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18841-95 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Related Products
Applications
Our Abpromise guarantee covers the use of ab208035 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 | 1/2000. Detects a band of approximately 46-54 kDa (predicted molecular weight: 48, 53 kDa). | |
ICC/IF | 1/1000. | |
Flow Cyt | 1/100. | |
IP | 1/40. |
Target
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Function
Serine/threonine-protein kinase involved in various processes such as cell proliferation, differentiation, migration, transformation and programmed cell death. Extracellular stimuli such as proinflammatory cytokines or physical stress stimulate the stress-activated protein kinase/c-Jun N-terminal kinase (SAP/JNK) signaling pathway. In this cascade, two dual specificity kinases MAP2K4/MKK4 and MAP2K7/MKK7 phosphorylate and activate MAPK8/JNK1. In turn, MAPK8/JNK1 phosphorylates a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 and ATF2 and thus regulates AP-1 transcriptional activity. Phosphorylates the replication licensing factor CDT1, inhibiting the interaction between CDT1 and the histone H4 acetylase HBO1 to replication origins. Loss of this interaction abrogates the acetylation required for replication initiation. Promotes stressed cell apoptosis by phosphorylating key regulatory factors including p53/TP53 and Yes-associates protein YAP1. In T-cells, MAPK8 and MAPK9 are required for polarized differentiation of T-helper cells into Th1 cells. Contributes to the survival of erythroid cells by phosphorylating the antagonist of cell death BAD upon EPO stimulation. Mediates starvation-induced BCL2 phosphorylation, BCL2 dissociation from BECN1, and thus activation of autophagy. Phosphorylates STMN2 and hence regulates microtubule dynamics, controlling neurite elongation in cortical neurons. In the developing brain, through its cytoplasmic activity on STMN2, negatively regulates the rate of exit from multipolar stage and of radial migration from the ventricular zone. Phosphorylates several other substrates including heat shock factor protein 4 (HSF4), the deacetylase SIRT1, ELK1, or the E3 ligase ITCH.
JNK1 isoforms display different binding patterns: beta-1 preferentially binds to c-Jun, whereas alpha-1, alpha-2, and beta-2 have a similar low level of binding to both c-Jun or ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms. -
Sequence similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. -
Domain
The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. -
Post-translational
modificationsDually phosphorylated on Thr-183 and Tyr-185 by MAP2K7 and MAP2K4, which activates the enzyme. Phosphorylated by TAOK2. -
Cellular localization
Cytoplasm. Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 5599 Human
- Entrez Gene: 5601 Human
- Entrez Gene: 5602 Human
- Entrez Gene: 26414 Mouse
- Entrez Gene: 26419 Mouse
- Entrez Gene: 26420 Mouse
- Entrez Gene: 116554 Rat
- Entrez Gene: 25272 Rat
see all -
Alternative names
- C Jun kinase 2 antibody
- c Jun N terminal kinase 1 antibody
- c Jun N terminal kinase 2 antibody
see all
Images
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All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution
Lane 1 : Human JNK1 full length recombinant protein
Lane 2 : Human JNK2 full length recombinant protein
Lane 3 : Human JNK3 full length recombinant protein
Lysates/proteins at 0.01 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 48, 53 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1 & 2: 5 seconds; Lane 3: 15 seconds.
Human JNK1 full length recombinant protein contains aa1-427 with a His-Tag®. Human JNK2 full length recombinant protein contains aa1-424 with a His-Tag®. Human JNK3 full length recombinant protein contains aa1-464 with a His-Tag®.
All three recombinant human full length proteins were made in-house.
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All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution
Lane 1 : Human fetal liver lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 48, 53 kDa
Observed band size: 46-54 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 48, 53 kDa
Observed band size: 46-54 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat kidney lysate
Lane 7 : 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
Predicted band size: 48, 53 kDa
Observed band size: 46-54 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 4 seconds; Lane 2,3 & 4: 30 seconds; Lane 5: 15 seconds; Lane 6 & 7: 3 minutes.
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All lanes : Anti-JNK1+JNK2+JNK3 antibody [EPR18841-95] (ab208035) at 1/2000 dilution
Lane 1 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 2 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 48, 53 kDa
Observed band size: 46-54 kDa why is the actual band size different from the predicted?
Exposure time: 8 secondsBlocking and Diluting buffer and concentration: 5% NFDM/TBST
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasm and weak nuclear staining on HeLa cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab208035 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing cytoplasm and nuclear staining on NIH/3T3 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab208035 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.
-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
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Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cell line from peripheral blood) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
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Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line labeling JNK1+JNK2+JNK3 with ab208035 at 1/100 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat anti-Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.
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JNK1+JNK2+JNK3 was immunoprecipitated from 1mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab208035 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab208035 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa whole cell lysate, 10ug (Input).
Lane 2: ab208035 IP in HeLa whole cell lysate.
Lane 3: Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab208035 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Protocols
Datasheets and documents
Certificate of Compliance
References (33)
ab208035 has been referenced in 33 publications.
- Hao Y et al. 2-Methylquinazoline derivative 23BB as a highly selective histone deacetylase 6 inhibitor alleviated cisplatin-induced acute kidney injury. Biosci Rep 40:N/A (2020). PubMed: 31894849
- Ma JX et al. Couplet medicines of leech and centipede granules improve erectile dysfunction via inactivation of the CaSR/PLC/PKC signaling in streptozotocin-induced diabetic rats. Biosci Rep 40:N/A (2020). PubMed: 31922200
- Xiang Y et al. miR-142-5p as a CXCR4-Targeted MicroRNA Attenuates SDF-1-Induced Chondrocyte Apoptosis and Cartilage Degradation via Inactivating MAPK Signaling Pathway. Biochem Res Int 2020:4508108 (2020). PubMed: 32047668
- Zhang Q et al. G6PD facilitates clear cell renal cell carcinoma invasion by enhancing MMP2 expression through ROS-MAPK axis pathway. Int J Oncol 57:197-212 (2020). PubMed: 32319593
- Zhao TM et al. Bicyclol Attenuates Acute Liver Injury by Activating Autophagy, Anti-Oxidative and Anti-Inflammatory Capabilities in Mice. Front Pharmacol 11:463 (2020). PubMed: 32362825