Recombinant Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5693] to JNK1 + JNK2 + JNK3 (phospho T183+T183+T221)
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, Dot blot
- Reacts with: Mouse, Human
Related conjugates and formulations
Overview
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Product name
Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693]
See all JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) primary antibodies -
Description
Rabbit monoclonal [EPR5693] to JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) -
Host species
Rabbit -
Specificity
ab124956 will detect will detect JNK1 (pT183), JNK2 (pT183) and JNK3 (pT221). -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF, Dot blotmore details -
Species reactivity
Reacts with: Mouse, Human
Predicted to work with: Rat -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- NIH 3T3 cell lysates treated with Anisomycin; Human brain tissue. IP: HeLa treated with 25ug/mL anisomycin for 30min 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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 2.09 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5693 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab201862)
- Alexa Fluor® 647 Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab201864)
- PE Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab208843)
- Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] - BSA and Azide free (ab219584)
- Alexa Fluor® 568 Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab312926)
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab124956 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 | (5) |
1/1000 - 1/10000. Detects a band of approximately 46-54 kDa.
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IP |
1/10 - 1/100.
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
(Heat to 98°C, allow to cool for 10-20 minutes) |
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ICC/IF | (1) |
1/50 - 1/100.
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Dot blot |
1/1000.
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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
1/1000 - 1/10000. Detects a band of approximately 46-54 kDa. |
IP
1/10 - 1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (Heat to 98°C, allow to cool for 10-20 minutes) |
ICC/IF
1/50 - 1/100. |
Dot blot
1/1000. |
Target
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Cellular localization
Cytoplasmic, Mitochondrial, Nuclear and Plasma membrane -
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
- JNK 46 antibody
- JNK 55 antibody
- MAPK10 antibody
see all
Images
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All lanes : Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates with 5% NFDM/TBST
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates with 5% NFDM/TBST
Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates. Then the membrane was incubated with alkaline phosphatase with 5% NFDM/TBST
Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates. Then the membrane was incubated with lambda phosphatase with 5% NFDM/TBST
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Observed band size: 46,54 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)
ab124956, at 1/100 dilution staining JNK1+JNK2+JNK3 in paraffin-embedded Human brain tissue, by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)
Immunocytochemistry/Immunofluorescence analysis of untreated, Anisomycin treated and Anisomycin + LP treated NIH/3T3 cells labelling JNK1 + JNK2 + JNK3 (phospho T183 + T183 + T221) with ab124956 at a dilution of 1/100 (left) and JNK1 + JNK2 + JNK3 with ab179461 at a dilution of 1/250 (right).
Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
The image shows increased nuclear staining after Anisomycin (250ng/ml, 30min) treatment on NIH3T3 cells. The LP treatment decreased the increased nuclear staining caused by Anisomycin.
ab179461 was used as a Pan control for ab124956. The results showed cytoplasmic staining on untreated, Anisomycin and Anisomycin + LP treated NIH3T3 cells.
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Flow Cytometry (Intracellular) - Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)
Overlay histogram showing HeLa cells stained with ab124956 (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 (ab124956, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluorr® 488 IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Immunoprecipitation - Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956)
Purified ab124956 at 1/70 dilution (2µg) immunoprecipitating JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) in HeLa treated with 25ug/mL anisomycin for 30min whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) treated with 25ug/mL anisomycin for 30min whole cell lysate 10µg
Lane 2 (+): ab124956 + HeLa treated with 25ug/mL anisomycin for 30min whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124956 in HeLa treated with 25ug/mL anisomycin for 30min whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 46, 54 kDa -
All lanes : Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (ab124956) at 1/1000 dilution
Lane 1 : NIH 3T3 cell lysate, untreated
Lane 2 : NIH 3T3 cell lysate, treated with Anisomycin
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilutionSecondary antibody - goat anti-rabbit HRP (ab6721)
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Dot blot analysis of JNK1/2/3 (pT183 + pT183 + pT221) peptide (Lane 1) and JNK1/2/3 non-phospho peptide (Lane 2) labelling JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) with ab124956 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (207)
ab124956 has been referenced in 207 publications.
- Chen Y et al. PP1A prevents ROS-induced pyroptosis by inhibiting MAPK/caspase-3 in mouse adipose tissue. FEBS J 289:3839-3853 (2022). PubMed: 35080339
- Wang C & Cheng B MicroRNA miR-3646 promotes malignancy of lung adenocarcinoma cells by suppressing sorbin and SH3 domain-containing protein 1 via the c-Jun NH2-terminal kinase signaling pathway. Bioengineered 13:4869-4884 (2022). PubMed: 35196185
- Huang Y et al. Butorphanol reduces the neuronal inflammatory response and apoptosis via inhibition of p38/JNK/ATF2/p53 signaling. Exp Ther Med 23:229 (2022). PubMed: 35222706
- Huang Z et al. MiR-146a alleviates lung injury caused by RSV infection in young rats by targeting TRAF-6 and regulating JNK/ERKMAPK signaling pathways. Sci Rep 12:3481 (2022). PubMed: 35241728
- Luo G et al. Blocking GSDME-mediated pyroptosis in renal tubular epithelial cells alleviates disease activity in lupus mice. Cell Death Discov 8:113 (2022). PubMed: 35279675