Phospho-JNK1/2/3 (T183 + T183 + T221) Recombinant Rabbit Monoclonal Antibody [ST500] - BSA and Azide free
Catalog# HA750173
Phospho-JNK1/2/3 (T183 + T183 + T221) Recombinant Rabbit Monoclonal Antibody [ST500] - BSA and Azide free
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WB
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IF-Cell
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IF-Tissue
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IHC-P
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IP
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FC
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IHC-Fr
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Human
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Mouse
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Rat
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ET1609-42
含抗保成分
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unconjugated
Overview
Product Name
Phospho-JNK1/2/3 (T183 + T183 + T221) Recombinant Rabbit Monoclonal Antibody [ST500] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Thr183 of Human JNK1 aa 161-204 / 427.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC, IHC-Fr
Molecular Weight
Predicted band size: 48/53 kDa
Positive Control
A431 cell lysate treated with UV40, 293 cell lysate treated with UV40, Hela cell lysate treated with UV40, HeLa treated with 25μg/mL Anisomycin for 30 minutes cell lysate, NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes cell lysate, C6 treated with 25ug/mL Anisomycin for 30 minutes cell lysate, Hela, NIH/3T3, HUVEC, human colon tissue, human endometrium tissue, mouse heart tissue, rat liver tissue, rat kidney tissue.
Conjugation
unconjugated
Clone Number
ST500
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:2,000-1:5,000
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IF-Cell
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1:100-1:200
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IF-Tissue
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1:200-1:500
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IHC-P
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1:400-1:1,000
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FC
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1:500-1:1,000
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IP
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Use at an assay dependent concentration.
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IHC-Fr
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1:100-1:500
Target
Function
JNKs (c-Jun N-terminal kinases) belong to a family of MAP kinases that are involved in a variety of cellular processes, including transcriptional regulation and cellular proliferation, differentiation and development. JNK2 (c-Jun N-terminal kinase 2) and JNK3 (c-Jun N-terminal kinase 3) are 424 and 464 amino acid proteins, respectively, that each contain one protein kinase domain and use magnesium as a cofactor to catalyze the phosphorylation of target proteins, thereby playing a role in a variety of events throughout the cell. Both JNK2 and JNK3 exist as multiple alternatively spliced isoforms and are subject to post-translational phosphorylation on Thr 183 and Thr 221, respectively, an event which activates JNK2/JNK3 enzymatic activity. Defects in the gene encoding JNK3 are a cause of epileptic encephalopathy of the Lennox-Gastaut type, a group of epileptic disorders characterized by severe psychomotor delay and seizures.
Background References
1. Kang K et al. Carnosic acid slows photoreceptor degeneration in the Pde6b(rd10) mouse model of retinitis pigmentosa. Sci Rep 6:22632 (2016).
2. Li C et al. Inhibitory effects of kaempferol on the invasion of human breast carcinoma cells by downregulating the expression and activity of matrix metalloproteinase-9. Biochem Cell Biol 93:16-27 (2015).
Sequence Similarity
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Post-translational Modification
Dually phosphorylated on Thr-183 and Tyr-185 by MAP2K7 and MAP2K4, which activates the enzyme. Phosphorylated by TAOK2. May be phosphorylated at Thr-183 and Tyr-185 by MAP3K1/MEKK1. Phosphorylated form is more concentrated at synapses than none-phosphorylated (By similarity).
Subcellular Location
Cytoplasm, Membrane, Mitochondrion, Nucleus.
UNIPROT
Synonyms
C Jun kinase 2 antibody
c Jun N terminal kinase 1 antibody
c Jun N terminal kinase 2 antibody
c Jun N terminal kinase 3 antibody
c-Jun N-terminal kinase 1 antibody
JNK 46 antibody
JNK 55 antibody
JNK antibody
JNK-46 antibody
JNK1 antibody
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/1,000 dilution.
Lane 1: Hela cell lysate, untreated (10 µg/Lane)
Lane 2: Hela cell lysate, treated with Anisomycin (10 µg/Lane)
Lane 3: A431 cell lysate, untreated (10 µg/Lane)
Lane 4: A431 cell lysate, treated with UV40 (10 µg/Lane)
Lane 5: 293 cell lysate, untreated (10 µg/Lane)
Lane 6: 293 cell lysate, treated with UV40 (10 µg/Lane)
Lane 7: Hela cell lysate, untreated (10 µg/Lane)
Lane 8: Hela cell lysate, treated with UV40 (10 µg/Lane)
Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa
Exposure time: 2 minutes;
12% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750173) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/2,000 dilution.
Lane 1: C6 cell lysate
Lane 2: C6 treated with 25µg/mL Anisomycin for 30 minutes cell lysate.
Lysates/proteins at 20 µg/Lane.
Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa
Exposure time: 2 minutes 6 seconds; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750173) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Western blot analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) on different lysates with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 25μg/mL Anisomycin for 30 minutes cell lysate
Lane 3: NIH/3T3 cell lysate
Lane 4: NIH/3T3 treated with 25μg/mL Anisomycin for 30 minutes cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 48/53 kDa
Observed band size: 48/53 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750173) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in Hela cells (green).
Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750173, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in NIH/3T3 cells (green).
Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750173, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
ICC staining of Phospho-JNK1/2/3 (T183 + T183 + T221) in HUVEC cells (green).
Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (HA750173, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue). -
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750173) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human endometrium tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750173) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/400 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750173) at 1/400 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750173) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750173) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of Phospho-JNK1/2/3 (T183 + T183 + T221) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (HA750173, 1/100) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
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Immunofluorescence analysis of frozen mouse hippocampus tissue labeling Phospho-JNK1/2/3 (T183 + T183 + T221) with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (HA750173, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner. -
Immunofluorescence analysis of frozen mouse cerebral cortex tissue labeling Phospho-JNK1/2/3 (T183 + T183 + T221) with Rabbit anti-Phospho-JNK1/2/3 (T183 + T183 + T221) antibody (HA750173).
The tissues were blocked in 3% BSA for 30 minutes at room temperature, washed with PBS, and then probed with the primary antibody (HA750173, green) at 1/100 dilution overnight at 4℃, washed with PBS. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) was used as the secondary antibody at 1/200 dilution. Nuclei were counterstained with DAPI (blue). Image acquisition was performed with KFBIO KF-FL-400 Scanner.
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