Anti-JNK2 抗体 [EP1595Y] (ab76125)
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
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ELISA
- Knockout validated
- Reacts with: Human, Recombinant fragment
Related conjugates and formulations
製品の概要
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製品名
Anti-JNK2 antibody [EP1595Y]
JNK2 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1595Y] to JNK2 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IP, IHC-P, ELISAmore details -
種交差性
交差種: Human, Recombinant fragment
交差が予測される動物種: Mouse, Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: HEK293T, MCF7, HAP1 and HeLa cell lysates. IHC-P: Human breast carcinoma tissue. IP: HeLa cell lysate. Flow Cyt (intra): HeLa cells.
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特記事項
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.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP1595Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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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
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ab76125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 |
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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000 - 1/10000. Predicted molecular weight: 48 kDa.
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IP |
Use a concentration of 5 µg/ml.
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IHC-P |
1/100 - 1/250. 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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特記事項 |
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Flow Cyt (Intra)
1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Predicted molecular weight: 48 kDa. |
IP
Use a concentration of 5 µg/ml. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ELISA
Use at an assay dependent concentration. |
ターゲット情報
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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 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. -
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. Autophosphorylated in vitro. - Information by UniProt
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参照データベース
- 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 -
別名
- c Jun kinase 2 antibody
- C Jun N terminal kinase 2 antibody
- c-Jun N-terminal kinase 2 antibody
see all
画像
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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 kDaLanes 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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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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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
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Anti-JNK2 antibody [EP1595Y] (ab76125) at 1/50000 dilution + HeLa cell lysate at 10 µg
Secondary
goat anti-rabbit-HRP at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 48 kDa
Observed band size: 54 kDa why is the actual band size different from the predicted?
Additional bands at: 46 kDa (possible isoform) -
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.
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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.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (39)
ab76125 は 39 報の論文で使用されています。
- Tian X et al. c-Jun N-terminal kinase 2 suppresses pancreatic cancer growth and invasion and is opposed by c-Jun N-terminal kinase 1. Cancer Gene Ther 29:73-86 (2022). PubMed: 33526844
- Dong W et al. Wnt4 is crucial for cardiac repair by regulating mesenchymal-endothelial transition via the phospho-JNK/JNK. Theranostics 12:4110-4126 (2022). PubMed: 35673578
- Gong M et al. PHLDA1 knockdown inhibits inflammation and oxidative stress by regulating JNK/ERK pathway, and plays a protective role in sepsis-induced acute kidney injury. Allergol Immunopathol (Madr) 50:1-9 (2022). PubMed: 36335439
- Yu W et al. TRPV3 inhibits colorectal cancer cell proliferation and migration by regulating the MAPK signaling pathway. J Gastrointest Oncol 13:2447-2457 (2022). PubMed: 36388700
- Zhang J et al. LncRNA SNHG14 contributes to proinflammatory cytokine production in rheumatoid arthritis via the regulation of the miR-17-5p/MINK1-JNK pathway. Environ Toxicol 36:2484-2492 (2021). PubMed: 34529319