概述

  • 产品名称Anti-MEKK2抗体[EP626Y]
    参阅全部 MEKK2 一抗
  • 描述
    兔单克隆抗体[EP626Y] to MEKK2
  • 经测试应用适用于: IHC-Fr, ICC/IF, WB, IHC-P, IP, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide. within Human MEKK2 aa 1-100 (N terminal). The exact sequence is proprietary.
    Database link: Q9Y2U5

  • 阳性对照
    • WB and Flow Cyt: Jurkat cell lysate, human breast carcinoma. IHC-P: Human colon carcinoma, mouse and rat cerebral cortex. ICCIF: MCF-7 cell line. IP: HepG2 cell line.
  • 常规说明

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • 存储溶液pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • 纯度Protein A purified
  • 克隆单克隆
  • 克隆编号EP626Y
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab33918 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
IHC-Fr Use at an assay dependent concentration.
ICC/IF 1/250 - 1/500.
WB 1/10000 - 1/50000. Predicted molecular weight: 70 kDa.
IHC-P 1/100.
IP 1/40.
Flow Cyt 1/50 - 1/120.

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. Regulates the JNK and ERK5 pathways by phosphorylating and activating MAP2K5 and MAP2K7 (By similarity). Plays a role in caveolae kiss-and-run dynamics.
  • 序列相似性Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
    Contains 1 OPR domain.
    Contains 1 protein kinase domain.
  • 翻译后修饰Autophosphorylated.
  • 细胞定位Cytoplasm. Nucleus. Upon EGF stimulation, translocates into the nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • M3K2_HUMAN antibody
    • Map3k2 antibody
    • MAPK/ERK kinase kinase 2 antibody
    • MEK kinase 2 antibody
    • MEKK 2 antibody
    • MEKK2b antibody
    • Mitogen activated protein kinase kinase kinase 2 antibody
    • Mitogen-activated protein kinase kinase kinase 2 antibody
    see all

Anti-MEKK2 antibody [EP626Y] 图像



  • Predicted band size : 70 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MEKK2 knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Human breast carcinoma lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab33918 observed at 75 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab33918 was shown to recognize MEKK2 when MEKK2 knockout samples were used, along with additional cross-reactive bands. Wild-type and MEKK2 knockout samples were subjected to SDS-PAGE. ab33918 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • ab33918 staining MEKK2 in human colon carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

     

  • ab33918 staining MEKK2 in MCF-7 (human breast carcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab7291 and ab150120 were used as counterstains for primary antibody ab75748 and secondary antibody ab150077 respectively and DAPI was used as a nuclear counterstain.

    Negative control 1: Rabbit primary antibody and anti-mouse secondary antibody (ab150120)
    Negative control 2: Mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077)

     

  • ab33918 staining MEKK2 in Jurkat (human acute T cell leukemia) cells by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/120. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/500 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

     

  • ab33918 immunoprecipitating MEKK2. 10µg of cell lysate was incubated with primary antibody at a dilution of 1/40 and VeriBlot for IP secondary antibody (HRP) (ab131366) at a dilution of 1/10000.

    Lane 1: HepG2 (human hepatocellular carcinoma) whole cell lysate (10ug)
    Lane 2: HepG2 (human hepatocellular carcinoma) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab33918 in HepG2 (human hepatocellular carcinoma) whole cell lysate

     

  • All lanes : Anti-MEKK2 antibody [EP626Y] (ab33918) at 1/20000 dilution

    Lane 1 : C6 (rat glioma) whole cell lysate
    Lane 2 : NIH/3T3 (mouse embryo) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size : 70 kDa
  • ab33918 staining MEKK2 in mouse cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • Overlay histogram showing HepG2 cells stained with unpurified ab33918 (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 (ab33918, 1/50 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • MEKK2 was immunoprecipitated using 0.5mg HepG2 whole cell extract, 10µg of unpurified Rabbit monoclonal [EP626Y] to MEKK2 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, HepG2 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 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab33918.
    Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) .
    Band: 75kDa: MEKK2.

  • All lanes : Anti-MEKK2 antibody [EP626Y] (ab33918) at 1/20000 dilution

    Lane 1 : Jurkat (human acute T cell leukemia) whole cell lysate
    Lane 2 : HeLa (human cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size : 70 kDa
    Additional bands at : 70 kDa. We are unsure as to the identity of these extra bands.
  • ab33918 staining MEKK2 in rat cerebral cortex tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500.

    Negative control 1: PBS in place of primary antibody.

  • + K562 (human chronic myelogenous leukemia) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 dilution

    Predicted band size : 70 kDa

Anti-MEKK2 antibody [EP626Y] (ab33918)参考文献

This product has been referenced in:
  • Mazur PK  et al. SMYD3 links lysine methylation of MAP3K2 to Ras-driven cancer. Nature 510:283-7 (2014). WB ; Mouse, Human . Read more (PubMed: 24847881) »
  • Brown RD  et al. MAP kinase kinase kinase-2 (MEKK2) regulates hypertrophic remodeling of the right ventricle in hypoxia-induced pulmonary hypertension. Am J Physiol Heart Circ Physiol 304:H269-81 (2013). Read more (PubMed: 23125215) »

See all 5 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10)
Sample Mouse Cell lysate - whole cell (lung cancer line LKR10)
Specification lung cancer line LKR10
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 22 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (8%)
Sample Human Cell lysate - nuclear (lung cancer line A549)
Specification lung cancer line A549
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Jul 22 2014

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