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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
  • 宿主
    Rabbit
  • 经测试应用
    适用于: WB, IHC-P, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Human
  • 免疫原

    Synthetic peptide corresponding to Human ASK1 aa 1-100 (N terminal).
    (Peptide available as ab224650)

  • 阳性对照
    • Human lung carcinoma tisuue, HeLa cell lysate, HeLa cells, RAW 264.7 whole cell lysates, Human pancreas, Mouse kidney.
  • 常规说明

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

     

    This product is a recombinant rabbit monoclonal antibody.

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 1/1000. Detects a band of approximately 155 kDa (predicted molecular weight: 155 kDa).Can be blocked with ASK1 peptide (ab224650).

For unpurified use at 1/2000

IHC-P 1/50. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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.

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

图片

  • 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.

    Unpurified 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence analysis of RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling ASK1 with Purified ab45178 at 1:100 (9.9 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 �g/ml). Goat anti rabbit IgG (Alexa Fluor� 488, ab150077) was used as the secondary antibody at 1:1000 (2 �g/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ASK1 with purified ab45178 at 1:100 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
  • Immunofluorescent staining of HeLa cells using unpurified ab45178 (1/10).

  • Overlay histogram showing HeLa cells stained with unpurified 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.

  • Unpurified ab45178 (1/10) staining human ASK1 in human lung carcinoma by immunohistochemistry using paraffin embedded tissue.

  • Anti-ASK1 antibody [EP553Y] (ab45178) at 1/2000 dilution (Unpurified) + 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

文献

This product has been referenced in:

See all 14 Publications for this product

客户评价及客户问答

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...

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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
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"

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