Anti-JNK1+JNK2 (phospho T183 + Y185)抗体(ab4821)

概述

  • 产品名称Anti-JNK1+JNK2 (phospho T183 + Y185)抗体
    参阅全部 JNK1+JNK2 一抗
  • 描述
    兔多克隆抗体to JNK1+JNK2 (phospho T183 + Y185)
  • 特异性Phosphorylation site-specific antibody selective for the dually phosphorylated form of the c-Jun N-terminal Kinase (JNK)/Stress-Activated Protein Kinase (SAPK) enzymes containing a phosphate on threonine 183 and tyrosine 185 (human JNK 1 + 2). The antibody has been shown to recognize the endogenous, active forms of JNK 1 + 2 in a variety of cell types following treatment by a broad range of extracellular stimuli [e.g. including 293 cells (human embryonic kidney; +/- ultraviolet light) and PC12 cells (rat pheochromocytoma; +/- sorbital)]. The region of JNK1 and JNK2 surrounding T183 + Y185 has a high degree of similarity to the corresponding regions in JNK3 and thus may cross react with this protein if phosphorylated on the corresponding residues.
  • 经测试应用适用于: ICC/IF, IHC-P, IHC-Fr, WBmore details
  • 种属反应性
    与反应: Mouse, Human
    预测可用于: a wide range of other species
  • 免疫原

    Synthetic peptide (Human) derived from the region of JNK 1 + 2 that contains threonine 183 and tyrosine 185, based on the human sequence. This region is conserved among many species including human, rat, mouse, chick, nematode (Caenorhabditis elegans), and fly (Drosophila melanogaster).

性能

应用

Our Abpromise guarantee covers the use of ab4821 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/1 - 1/100. 1/100.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 49, 55 kDa.

Band at ~49 kDa represents Jnk1, while the band at ~55 kDa represents Jnk2

靶标

  • 功能Responds to activation by environmental stress and pro-inflammatory cytokines by phosphorylating a number of transcription factors, primarily components of AP-1 such as JUN, JDP2 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 (By similarity). Phosphorylates heat shock factor protein 4 (HSF4).
    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.
  • 序列相似性Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • 结构域The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • 翻译后修饰Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme.
  • Information by UniProt
  • 数据库链接
  • 别名
    • c jun N terminal kinase 2 antibody
    • c-Jun N-terminal kinase 1 antibody
    • cJun N terminal kinase 1 antibody
    • JNK 1 antibody
    • JNK 2 antibody
    • JNK-46 antibody
    • JNK2ALPHA antibody
    • JNK2BETA antibody
    • MAP kinase 8 antibody
    • MAP kinase 9 antibody
    • MAPK 8 antibody
    • mapk8 antibody
    • MAPK9 antibody
    • Mitogen activated protein kinase 8 antibody
    • Mitogen activated protein kinase 9 antibody
    • Mitogen-activated protein kinase 8 antibody
    • MK08_HUMAN antibody
    • PRKM 8 antibody
    • PRKM 9 antibody
    • Stress-activated protein kinase 1 antibody
    • Stress-activated protein kinase JNK1 antibody
    see all

Anti-JNK1+JNK2 (phospho T183 + Y185) antibody 图像


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 49, 55 kDa


    Exposure time : 20 minutes

    MEF1 cells were incubated at 37°C for 48h with vehicle control (0 µM) and 5 µM of glibenclamide (ab120267) in DMSO. Increased expression of of JNK1+JNK2 (phospho T183 + Y185) (ab4821) correlates with an increase in glibenclamide concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 3% milk before being incubated with ab4821 at 1/1000 dilution and ab85139 at 1 µg /ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.



  • Predicted band size : 49, 55 kDa
    To demonstrate the phosphorylation of JNK 1 & 2 in a cell based assay, 293 cells were treated with ultraviolet irradiation (UV). Proteins from cell extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were incubated with either 1 µ g/mL ab4821 or 1 µg/mL anti-JNK1 pan. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix WesternStar detection method.
    To demonstrate the phosphorylation of JNK 1 & 2 in a cell based assay, 293 cells were treated with ultraviolet irradiation (UV). Proteins from cell extracts were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to nitrocellulose. Membranes were incubated with either 1 µ g/mL ab4821 or 1 µg/mL anti-JNK1 pan. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and bands were detected using the Tropix We


  • Predicted band size : 49, 55 kDa

    MCF7cells were incubated at 37°C for 4h with vehicle control (0 µM) and different concentrations of cryptotanshinone (ab120666). Increased expression of JNK1+JNK2 (phospho T183 + Y185) in MCF7 cells correlates with an increase in cryptotanshinone concentration, as described in literature.

    Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab4821 at 1/1000 dilution and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP (ab97051) at 1/10000 dilution and visualised using ECL development solution.

  • Figure shows immunostaining of fibroblast podosomes using ab4821. Green = dual phospho JNK, Red = mitochondria, Blue = nucleus.
  • ab4821 staining JNK1+JNK2 (phospho T183 + Y185) in human foreskin fibroblasts by ICC/IF. The cells were fixed in cytoskeletal fixative, permeabilized in 0.5% Triton X-100 and blocked in 2% dillution buffer (2%BSA + 0.1% Triton X-100) for 1 hour at 25°C. The primary antibody was diluted, 1/100 and incubated with sample for 12 hours. An Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG, diluted 1/250 was used as secondary.

    See Abreview

  • ab4821 staining JNK1+JNK2 (phospho T183 + Y185) in E12.5 mouse heart tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent cold 1% paraformaldehyde fixation before heat mediated antigen retrieval with Tris-EDTA pH9 and then blocking with 2.25% horse serum was performed for 20 minutes at RT. The primary antibody was diluted 1/400 and incubated with sample for 12 hours at 4°C. A Biotin conjugated horse polyclonal to rabbit IgG was used at dilution at 1/133 as secondary antibody.

    See Abreview

Anti-JNK1+JNK2 (phospho T183 + Y185) antibody (ab4821)参考文献

This product has been referenced in:
  • Li Q  et al. Chronic sciatic nerve compression induces fibrosis in dorsal root ganglia. Mol Med Rep 13:2393-400 (2016). Rat . Read more (PubMed: 26820076) »
  • Guo J  et al. Berberine Protects Human Umbilical Vein Endothelial Cells against LPS-Induced Apoptosis by Blocking JNK-Mediated Signaling. Evid Based Complement Alternat Med 2016:6983956 (2016). WB ; Human . Read more (PubMed: 27478481) »

See all 14 Publications for this product

Product Wall


When making quantitative comparisons, such as standardizing the levels of phospho-JNK to total JNK, we do not recommend stripping and reprobing the membrane, as stripping removes some sample protein from the membrane. Instead, you should either ru...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (myofibroblast stromal)
Loading amount 30 µg
Specification myofibroblast stromal
Gel Running Conditions Reduced Denaturing (gel 10 %)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Abcam user community

Verified customer

提交于 Apr 02 2013

Thank you very much for your interest our products.

To our knowledge, ab4821 has not been experimentally tested in rat. However by participating in our AbTrial program you can now use our products in an untested application or species withou...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Jurkat cells)
Loading amount 10 µg
Specification Jurkat cells
Treatment Untreated Jurkat cells and cells treated with 10-8, 10-7, and 10-6 uM Methotrexate for 48 hours
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 15°C
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Abcam user community

Verified customer

提交于 Sep 15 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (primary neurons)
Loading amount 30 µg
Specification primary neurons
Treatment 0.5M sorbitol
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Sep 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (used for heart and neural tube sections)
Specification used for heart and neural tube sections
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 2.25% · Temperature: RT°C
Username

Mrs. Michelle Combs

Verified customer

提交于 Oct 05 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Foreskin Fibroblast)
Specification Foreskin Fibroblast
Fixative Cytoskeletal Fixative
Permeabilization Yes - 0.5% Triton X-100
Blocking step Abdil (2%BSA + 0.1% Triton X-100) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
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Mr. George Chennell

Verified customer

提交于 Mar 03 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 30 µg
Specification HEK293
Treatment 150uM anisomycin for 30mins
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

提交于 Jul 01 2008

Thank you for your enquiry. The epitopes for both ab4821 and ab10664 are conserved for JNK1, 2 and 3. These antibodies will most likely cross react with JNK3. I hope this information helps, please do not hesitate to contact us if you need any more ...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa cells)
Loading amount 10 µg
Specification HeLa cells
Treatment 20ng/ml TNFSF2 for 0, 15, and 30min
Blocking step Milk as blocking agent for 20 minute(s) · Concentration: 3%
Username

Abcam user community

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提交于 Sep 27 2006

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