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RabMAb

Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) 抗体 [EPR5693] (ab124956)

製品の概要

  • 製品名
    Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693]
    JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR5693] to JNK1 + JNK2 + JNK3 (phospho T183+T183+T221)
  • 由来種
    Rabbit
  • 特異性
    ab124956 will detect will detect JNK1 (pT183), JNK2 (pT183) and JNK3 (pT221).
  • アプリケーション
    適用あり: Flow Cyt, WB, IP, IHC-P, ICC, ICC/IF, Dot blotmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human JNK1 + JNK2 + JNK3 (phospho T183+T183+T221).

  • ポジティブ・コントロール
    • NIH 3T3 cell lysates treated with Anisomycin; Human brain tissue.
  • 特記事項

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt 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 1/50 - 1/100.
ICC/IF Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.

ターゲット情報

画像

  • 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
  • ab124956, at 1/100 dilution staining JNK1+JNK2+JNK3 in paraffin-embedded Human brain tissue, by Immunohistochemistry.
  • 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.

  • 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 Fluor® 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.

  • 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 dilution


    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

参考文献

This product has been referenced in:
  • Zhang X  et al. Tanshinone IIA induces apoptosis of ovarian cancer cells in vitro and in vivo through attenuation of PI3K/AKT/JNK signaling pathways. Oncol Lett 17:1896-1902 (2019). Read more (PubMed: 30675253) »
  • Wang M  et al. miR-302a inhibits human HepG2 and SMMC-7721 cells proliferation and promotes apoptosis by targeting MAP3K2 and PBX3. Sci Rep 9:2032 (2019). Read more (PubMed: 30765768) »
See all 55 Publications for this product

レビューと Q&A

1-7 of 7 Abreviews or Q&A

Application
Western blot
Sample
Dog Cell lysate - whole cell (fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
fibroblasts
Blocking step
(agent) for 50 minute(s) · Concentration: 100%

Dr. 令 中野

Verified customer

投稿 Jun 20 2019

Application
Western blot
Sample
Rat Cell lysate - whole cell (Neruo cell)
Gel Running Conditions
Reduced Denaturing (12 %)
Loading amount
20 µg
Specification
Neruo cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 27°C

Miss. Ji Yeon Lee

Verified customer

投稿 Jan 25 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Human Glioblastoma cell line U87MG)
Permeabilization
Yes - PBST 0.1%
Specification
Human Glioblastoma cell line U87MG
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Mr. Matthias Dedobbeleer

Verified customer

投稿 Jun 08 2018

Application
Western blot
Sample
Mouse Tissue lysate - whole (retina)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
40 µg
Specification
retina
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%

Abcam user community

Verified customer

投稿 Jun 10 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 21°C
Sample
Mouse Tissue sections (retina)
Specification
retina
Permeabilization
Yes - 0.5% Triton X100 in PBS
Fixative
Paraformaldehyde

Abcam user community

Verified customer

投稿 Mar 10 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (HCT116 colon cancer cell line)
Loading amount
30 µg
Specification
HCT116 colon cancer cell line
Treatment
DMSO control and 10 µM Irinotecan for 16 hrs
Gel Running Conditions
Reduced Denaturing (13%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Christian Marx

Verified customer

投稿 Feb 22 2013

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of alternative ab124956with the order number ########.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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