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Anti-ASK1抗体[EP553Y] (ab45178)

概述

  • 产品名称Anti-ASK1抗体[EP553Y]
    参阅全部 ASK1 一抗
  • 描述
    兔单克隆抗体[EP553Y] to ASK1
  • 经测试应用适用于: WB, IHC-P, ICC/IF, Flow Cytmore details
    不适用于: IP
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide corresponding to residues in the N-terminus of human ASK1.

  • 阳性对照
    • Human lung carcinoma tisuue, HeLa cell lysate, HeLa cells.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

应用

Our Abpromise guarantee covers the use of ab45178 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
WB 1/2000. Detects a band of approximately 155 kDa (predicted molecular weight: 155 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
Flow Cyt 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

  • 应用说明Is unsuitable for IP.
  • 靶标

    • 功能Component of a protein kinase signal transduction cascade. Phosphorylates and activates MAP2K4 and MAP2K6, which in turn activate the JNK and p38 MAP kinases, respectively. Overexpression induces apoptotic cell death.
    • 组织特异性Abundantly expressed in heart and pancreas.
    • 序列相似性Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
      Contains 1 protein kinase domain.
    • 翻译后修饰Dephosphorylated and activated by PGAM5.
    • Information by UniProt
    • 数据库链接
    • 别名
      • Apoptosis signal regulating kinase 1 antibody
      • Apoptosis signal-regulating kinase 1 antibody
      • ASK 1 antibody
      • ASK-1 antibody
      • ASK1 antibody
      • M3K5 antibody
      • M3K5_HUMAN antibody
      • MAP/ERK kinase kinase 5 antibody
      • MAP3K5 antibody
      • MAPK/ERK kinase kinase 5 antibody
      • MAPKKK5 antibody
      • MEK kinase 5 antibody
      • MEKK 5 antibody
      • MEKK5 antibody
      • Mitogen activated protein kinase kinase kinase 5 antibody
      • Mitogen-activated protein kinase kinase kinase 5 antibody
      see all

    Anti-ASK1 antibody [EP553Y] 图像



    • Predicted band size : 155 kDa

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: ASK1 knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lane 4: A549 cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab45178 observed at 155, 160 kDa. Red - loading control, ab1240, observed at 124 kDa.

      ab45178 was shown to recognize ASK1 when ASK1 knockout samples were used, along with additional cross-reactive bands. Wild-type and ASK1 knockout samples were subjected to SDS-PAGE. ab45178 and ab1240 (loading control to Vinculin) were diluted to 1/2000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

    • Overlay histogram showing HeLa cells stained with ab45178 (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 (ab45178, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was a goat anti-rabbit DyLight® 488 (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.

    • Ab45178 (1/10) staining human ASK1 in human lung carcinoma by immunohistochemistry using paraffin embedded tissue.
    • Immunofluorescent staining of HeLa cells using ab45178 (1/10).
    • Anti-ASK1 antibody [EP553Y] (ab45178) at 1/2000 dilution + HeLa cell lysate at 10 µg

      Secondary
      Goat anti-rabbit, HRP labelled at 1/2000 dilution

      Predicted band size : 155 kDa
      Observed band size : 155 kDa

    Anti-ASK1 antibody [EP553Y] (ab45178)参考文献

    This product has been referenced in:
    • Maruyama T  et al. Roquin-2 promotes ubiquitin-mediated degradation of ASK1 to regulate stress responses. Sci Signal 7:ra8 (2014). Read more (PubMed: 24448648) »
    • Mosallanejad K  et al. The DEAH-box RNA helicase DHX15 activates NF-?B and MAPK signaling downstream of MAVS during antiviral responses. Sci Signal 7:ra40 (2014). Read more (PubMed: 24782566) »

    See all 9 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 25 µg
    Gel Running Conditions Reduced Denaturing
    Sample Mouse Cell lysate - whole cell (LIVER)
    Specification LIVER
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Abcam user community

    Verified customer

    提交于 Nov 10 2014

    Currently we do not offer ab45178 without BSA, however we may be able to make this custom formulation for you. I'll put you in touch with our RabMAb Technical Account Executive and he can help you look into this further.

    The concentration of ab45178, lot GR48990-3, is 1.262 mg/mL.

    I have CC's your request for a quote for 10 vials of this lot to our bulk order specialist, who can be reached at sales@abcam.com. She will be able to provide you a quote which incl...

    Read More

    Thanks for your enquiry.

    If you could send along the lot number Iwill be able to provide the antibody concentration.

    Thank you for your enquiry.

    I think the recommendation to use antigen retrieval listed for ICC/IFis a mistake. Antigen retrieval should not be necessary for ICC. Here is a link to a good general ICC protocol on our website if you need it: Read More

    Thank you for your enquiry.

    The rabbit monoclonal isotype control antibody that was used in this flow experiment is not available on our catalog. We do have a rabbit polyclonal isotype control, http://www.abcam.com/Rabbit-polyclonal-IgG-ab274...

    Read More

    Thank you for your enquiry.

    On our online datasheet for ab45178, there is a FACS figure with the accompanying figure legend:

    "Overlay histogram showing HeLa cells stained with ab45178 (red line). The cells were fixed with 80% m...

    Read More

    Thank you for your reply. My colleague Kate is currently out of the office. I would be happy to refund or replace this product for you as you are experiencing problems. I understand it can be frustrating to change the conditions to obtain optima...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (Bone-marrow derived dendritic cells)
    Loading amount 70 µg
    Specification Bone-marrow derived dendritic cells
    Gel Running Conditions Reduced Denaturing (8% SDS gel)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Caglar Cekic

    Verified customer

    提交于 May 13 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Mouse Cell lysate - whole cell (neuron)
    Loading amount 20 µg
    Specification neuron
    Gel Running Conditions Reduced Denaturing
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
    Username

    Abcam user community

    Verified customer

    提交于 Jun 03 2008

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"