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  1. Link

    jnk1--jnk2-phospho-t183--y185-antibody-ab4821.pdf

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Signal Transduction Protein Phosphorylation Ser / Thr Kinases MAPK Pathway
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Anti-JNK1 + JNK2 (phospho T183 + Y185) 抗体 (ab4821)

  • Datasheet
Reviews (9)Q&A (3)References (81)

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Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)

Key features and details

  • Rabbit polyclonal to JNK1 + JNK2 (phospho T183 + Y185)
  • Suitable for: ICC/IF, WB
  • Reacts with: Mouse, Human
  • Isotype: IgG

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関連製品

製品の概要

  • 製品名

    Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody
    JNK1 + JNK2 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to JNK1 + JNK2 (phospho T183 + Y185)
  • 由来種

    Rabbit
  • 特異性

    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.
  • アプリケーション

    適用あり: ICC/IF, WBmore details
  • 種交差性

    交差種: Mouse, Human
    交差が予測される動物種: a wide range of other species
  • 免疫原

    Synthetic peptide corresponding to Human JNK1 + JNK2 (phospho T183 + Y185).

  • 特記事項

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • バッファー

    pH: 7.30
    Preservative: 0.05% Sodium azide
    Constituents: PBS, 50% Glycerol, 0.1% BSA

    BSA is IgG and protease free
  • Concentration information loading...
  • 精製度

    Immunogen affinity purified
  • 特記事項(精製)

    Purified from rabbit serum by sequential epitope specific chromatography. The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated JNK enzymes. The final product is generated by affinity chromatography using a JNK-derived peptide that is phosphorylated at threonine 183 and tyrosine 185, within the activation loop. Note: It is the dually phosphorylated form of these enzymes that has full enzymatic activity.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • MAPK Pathway
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Proto-oncogenes
    • Cancer
    • Signal transduction
    • Protein phosphorylation
    • Serine/threonine kinases
    • MAPK pathway
    • Cancer
    • Oncoproteins/suppressors
    • Oncoproteins
    • Signal transducers
    • Immunology
    • Innate Immunity
    • TLR Signaling

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Glibenclamide (Glyburide), K+ channel blocker (ab120267)
    • Cryptotanshinone, STAT3 inhibitor (ab120666)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab4821の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
ICC/IF (1)
1/250.

1/100.

WB (7)
1/1000. Predicted molecular weight: 49, 55 kDa.

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

特記事項
ICC/IF
1/250.

1/100.

WB
1/1000. Predicted molecular weight: 49, 55 kDa.

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

ターゲット情報

  • 機能

    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.
  • 配列類似性

    Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
    Contains 1 protein kinase domain.
  • ドメイン

    The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases.
  • 翻訳後修飾

    Dually phosphorylated on Thr-183 and Tyr-185, which activates the enzyme.
  • Target information above from: UniProt accession P45983 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 5599 Human
    • Entrez Gene: 5601 Human
    • Entrez Gene: 26419 Mouse
    • Entrez Gene: 26420 Mouse
    • Omim: 601158 Human
    • Omim: 602896 Human
    • SwissProt: P45983 Human
    • SwissProt: P45984 Human
    • SwissProt: Q91Y86 Mouse
    • SwissProt: Q9WTU6 Mouse
    • Unigene: 138211 Human
    • Unigene: 21495 Mouse
    see all
  • 別名

    • 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

画像

  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    All lanes : Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821) at 1/1000 dilution

    Lane 1 : HEK-293 cell line
    Lane 2 : HEK-293 treated for 5 minutes with 200 mM of Anisomycin
    Lane 3 : HEK-293 treated for 20 minutes with UV
    Lane 4 : MCF7 cell line
    Lane 5 : MCF7 treated for 5 minutes with 200 mM of Anisomycin
    Lane 6 : K562 cell line
    Lane 7 : K562 treated for 20 minutes with UV
    Lane 8 : HeLa cell line
    Lane 9 : HeLa treated for 20 minutes with UV

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/5000 dilution

    Predicted band size: 49, 55 kDa



    Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature.

  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)

    ab4821 staining JNK1 + JNK2 (phospho T183 + Y185) in A549 cells (green, panel a) by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.25% Triton X-100 and blocked with 5% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (2ug/ml in 1% BSA) for 3 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/400). Nuclei stained with DAPI (blue, panel b), F-actin stained with Alexa Fluor® 594 Phalloidin (red, panel b) and merged images (panel d).

  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    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
  • Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Western blot - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)
    Immunocytochemistry/ Immunofluorescence - Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody (ab4821)This image is courtesy of an Abreview submitted by Mr George Chennell

    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

プロトコール

  • Peptide Competition Experiment
  • Western blotting protocol

Click here to view the general protocols

データシートおよび資料

  • Datasheet download

    Download

参考文献 (81)

ab4821 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab4821 は 81 報の論文で使用されています。

  • Chen Q  et al. Shenyan Kangfu tablet alleviates diabetic kidney disease through attenuating inflammation and modulating the gut microbiota. J Nat Med 75:84-98 (2021). PubMed: 32997272
  • Da Q  et al. TAK1 is involved in sodium L-lactate-stimulated p38 signaling and promotes apoptosis. Mol Cell Biochem 476:873-882 (2021). PubMed: 33111211
  • Jiao CY  et al. BUB1B promotes extrahepatic cholangiocarcinoma progression via JNK/c-Jun pathways. Cell Death Dis 12:63 (2021). PubMed: 33431813
  • Chang C  et al. Regulatory role of the TLR4/JNK signaling pathway in sepsis-induced myocardial dysfunction. Mol Med Rep 23:N/A (2021). PubMed: 33760172
  • Jung HY  et al. NOX1 Inhibition Attenuates Kidney Ischemia-Reperfusion Injury via Inhibition of ROS-Mediated ERK Signaling. Int J Mol Sci 21:N/A (2020). PubMed: 32967113
View all Publications for this product

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1-10 of 12 Abreviews or Q&A

Western blot abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Below Average
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Dog Cell lysate - whole cell (canine synovial fibroblasts)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
10 µg
Specification
canine synovial fibroblasts
Blocking step
Blockace as blocking agent for 50 minute(s) · Concentration: 100% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Apr 12 2022

Western blot abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (myofibroblast stromal)
Gel Running Conditions
Reduced Denaturing (gel 10 %)
Loading amount
30 µg
Specification
myofibroblast stromal
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

投稿 Apr 02 2013

Western blot abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkat cells)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Treatment
Untreated Jurkat cells and cells treated with 10-8, 10-7, and 10-6 uM Methotrexate for 48 hours
Specification
Jurkat cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 15°C
Read More

Abcam user community

Verified customer

投稿 Sep 15 2011

Western blot abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (primary neurons)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Treatment
0.5M sorbitol
Specification
primary neurons
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

投稿 Sep 14 2010

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (used for heart and neural tube sections)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH9
Permeabilization
No
Specification
used for heart and neural tube sections
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2.25% · Temperature: RT°C
Fixative
Paraformaldehyde
Read More

Mrs. Michelle Combs

Verified customer

投稿 Oct 05 2009

Immunocytochemistry/ Immunofluorescence abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Foreskin Fibroblast)
Permeabilization
Yes - 0.5% Triton X-100
Specification
Foreskin Fibroblast
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
Fixative
Cytoskeletal Fixative
Read More

Mr. George Chennell

Verified customer

投稿 Mar 03 2009

Western blot abreview for Anti-JNK1 + JNK2 (phospho T183 + Y185) antibody

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Treatment
150uM anisomycin for 30mins
Specification
HEK293
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Read More

Abcam user community

Verified customer

投稿 Jul 01 2008

Question

Hello, I have purchased this antibody (Ab4821) to quantify levels of phopsho-JNK 1/2 in human astrocyte lysates. I would like to standardize the phospho-JNK levels to total JNK (Ab112501). Would you suggest that I strip the membrane? If not, what other methods would be available?

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Abcam community

Verified customer

Asked on Oct 01 2013

Answer


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 run 2 separate blots or run 1 blot and cut it in half to stain with each antibody separately.

Read More

Kevin Hanson

Abcam Scientific Support

Answered on Oct 01 2013

Question

Hi,



I am planning to use this antibody (ab4821) in rat cell line. In the specificity section of the datasheet, it is written that it recognizes the active forms of JNK 1 + 2 in PC12 cells.



Do you have any scientific reference for that ?



Thanks in advance for your answer.



Best regards,

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Abcam community

Verified customer

Asked on Dec 19 2012

Answer

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 without financial risk.

Simply follow these easy steps below to apply for our AbTrial Program:

1. Reply to this email, letting us know you are interested in testing this product.

2. Our scientists will email you an inactive personal discount code for the value of the product.

3. Purchase and test the product at the regular price.

4. Submit your results, including your discount code in the additional notes section of your Abreview.

5. Once the Abreview is submitted, the discount code will become active.

6. Apply your discount code on your next order to receive that value off.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/abtrial.

Read More

Abcam Scientific Support

Answered on Dec 19 2012

Question

I need to order a specific antibody against JNK1 and JNK2 which does not cross-react with JNK3. The antibody I want (ab37228) is out of stock until July which is too long to wait and so I was looking at buying ab4821. However, it doesn't state whether this is specific for JNK1 & JNK2 without detecting JNK3 and when I look at specific JNK1 antibody ab10664 it seems that the Western blot in the datasheet for this is identical to that for ab4821 but with different labelling for lane B (apparently specific JNK1 antibody in one but a pan JNK1/JNK2 in the other).

Read More

Abcam community

Verified customer

Asked on Apr 24 2007

Answer

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 advice or information.

Read More

Abcam Scientific Support

Answered on Apr 24 2007

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製品・研究分野別
  • 癌
  • 循環器
  • 細胞生物学
  • 発生
  • エピジェネティクス&核内情報
  • 免疫学
  • メタボリズム
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