Recombinant Anti-JNK2 antibody [EP1595Y] - BSA and Azide free (ab227986)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1595Y] to JNK2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ELISA
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
Overview
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Product name
Anti-JNK2 antibody [EP1595Y] - BSA and Azide free
See all JNK2 primary antibodies -
Description
Rabbit monoclonal [EP1595Y] to JNK2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ELISAmore details -
Species reactivity
Reacts with: Human, Recombinant fragment
Predicted to work with: Mouse, Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HEK293T, MCF7, HAP1 and HeLa cell lysates. IP: HeLa cell lysate. Flow Cyt (intra): HeLa cells. IHC-P: Human breast carcinoma tissue.
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General notes
ab227986 is the carrier-free version of ab76125.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1595Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- PE Anti-JNK2 antibody [EP1595Y] (ab306259)
- APC Anti-JNK2 antibody [EP1595Y] (ab306260)
- HRP Anti-JNK2 antibody [EP1595Y] (ab306261)
- Alexa Fluor® 488 Anti-JNK2 antibody [EP1595Y] (ab309634)
- Alexa Fluor® 647 Anti-JNK2 antibody [EP1595Y] (ab310000)
- Alexa Fluor® 594 Anti-JNK2 antibody [EP1595Y] (ab310363)
- Alexa Fluor® 555 Anti-JNK2 antibody [EP1595Y] (ab311886)
- Alexa Fluor® 568 Anti-JNK2 antibody [EP1595Y] (ab312355)
- Anti-JNK2 antibody [EP1595Y] (ab76125)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab227986 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) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ELISA |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 48 kDa. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ELISA
Use at an assay dependent concentration. |
Target
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Function
Responds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating a number of transcription factors, primarily components of AP-1 such as c-Jun and ATF2 and thus regulates AP-1 transcriptional activity. In T-cells, JNK1 and JNK2 are required for polarized differentiation of T-helper cells into Th1 cells.
JNK2 isoforms display different binding patterns: alpha-1 and alpha-2 preferentially bind to c-Jun, whereas beta-1 and beta-2 bind to ATF2. However, there is no correlation between binding and phosphorylation, which is achieved at about the same efficiency by all isoforms. JUNB is not a substrate for JNK2 alpha-2, and JUND binds only weakly to it. -
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, which activates the enzyme. Autophosphorylated in vitro. - Information by UniProt
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Database links
- Entrez Gene: 5601 Human
- Entrez Gene: 26420 Mouse
- Entrez Gene: 50658 Rat
- Omim: 602896 Human
- SwissProt: P45984 Human
- SwissProt: Q9WTU6 Mouse
- SwissProt: P49186 Rat
- Unigene: 484371 Human
see all -
Alternative names
- c Jun kinase 2 antibody
- C Jun N terminal kinase 2 antibody
- c-Jun N-terminal kinase 2 antibody
see all
Images
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All lanes : Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : MAPK9 knockout HEK293T cell lysate
Lane 3 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 48 kDa
Observed band size: 48 kDaThis data was developed using the same antibody clone in a different buffer formulation (ab76125).
Lanes 1-3: Merged signal (red and green). Green - ab76125 observed at 48 kDa. Red - loading control ab8245 observed at 36 kDa.
ab76125 Anti-JNK2 antibody [EP1595Y] was shown to specifically react with JNK2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266355 (knockout cell lysate ab257527) was used. Wild-type and JNK2 knockout samples were subjected to SDS-PAGE. ab76125 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 20000 dilution for 1 hour at room temperature before imaging.
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This WB data was generated using the same anti-JNK2 antibody clone, EP1595Y, in a different buffer formulation (cat# ab76125).
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: JNK2 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: MCF7 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab76125 observed at 54 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab76125 was shown to specifically react with JNK2 when JNK2 knockout samples were used. Wild-type and JNK2 knockout samples were subjected to SDS-PAGE. ab76125 and ab8245 (loading control to GAPDH) were diluted 1/2500 and 1/2000 respectively and incubated overnight at 4°C. 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 1/10000 dilution for 1 h at room temperature before imaging. -
JNK2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit monoclonal to JNK2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab76125.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 48kDa; JNK2
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76125).
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Overlay histogram showing HeLa cells stained with ab76125 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76125, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76125).
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This IHC data was generated using the same anti-JNK2 antibody clone, EP1595Y, in a different buffer formulation (cat# ab76125).
ab76125 at 1/100 dilution staining JNK2 in human breast carcinoma by Immunohistochemistry, Paraffin-embedded tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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This data was developed using ab76125, the same antibody clone in a different buffer formulation.
ELISA analysis of Human JNK2 recombinant protein at 250 ng/mL with ab76125. An Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/2500 dilution was used as the secondary antibody.
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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Datasheet download
Certificate of Compliance
References (10)
ab227986 has been referenced in 10 publications.
- Kitanaka T et al. JNK activation is essential for activation of MEK/ERK signaling in IL-1ß-induced COX-2 expression in synovial fibroblasts. Sci Rep 7:39914 (2017). RT-PCR ; Cat . PubMed: 28054591
- Zhu Q et al. Cepharanthine exerts antitumor activity on choroidal melanoma by reactive oxygen species production and c-Jun N-terminal kinase activation. Oncol Lett 13:3760-3766 (2017). PubMed: 28529590
- Wang B et al. IL-34 Upregulated Th17 Production through Increased IL-6 Expression by Rheumatoid Fibroblast-Like Synoviocytes. Mediators Inflamm 2017:1567120 (2017). WB ; Human . PubMed: 28659662
- Luan Q et al. RIPK1 regulates survival of human melanoma cells upon endoplasmic reticulum stress through autophagy. Autophagy 11:975-94 (2015). WB ; Human . PubMed: 26018731
- Yan T et al. Luteolin inhibits behavioral sensitization by blocking methamphetamine-induced MAPK pathway activation in the caudate putamen in mice. PLoS One 9:e98981 (2014). WB ; Mouse . PubMed: 24901319
- Peng Y & Zhang L Activation of the TLR1/2 pathway induces the shaping of the immune response status of peripheral blood leukocytes. Exp Ther Med 7:1708-1712 (2014). WB ; Human . PubMed: 24926371
- Lee JJ et al. High-mobility group box 1 protein is implicated in advanced glycation end products-induced vascular endothelial growth factor A production in the rat retinal ganglion cell line RGC-5. Mol Vis 18:838-50 (2012). WB . PubMed: 22511847
- Ke H et al. The c-Jun NH2-terminal kinase 2 plays a dominant role in human epidermal neoplasia. Cancer Res 70:3080-8 (2010). WB, IP ; Human . PubMed: 20354187
- Zhao Y & Herdegen T Cerebral ischemia provokes a profound exchange of activated JNK isoforms in brain mitochondria. Mol Cell Neurosci 41:186-95 (2009). PubMed: 19289169
- Chaveroux C et al. Identification of a novel amino acid response pathway triggering ATF2 phosphorylation in mammals. Mol Cell Biol 29:6515-26 (2009). PubMed: 19822663