概述

  • 产品名称Anti-NEDD8抗体[Y297]
    参阅全部 NEDD8 一抗
  • 描述
    兔单克隆抗体[Y297] to NEDD8
  • 特异性ab81264 detects NEDD8 bound to cullins proteins (neddylated cullins) as well as free NEDD8.
  • 经测试应用适用于: WB, IHC-P, ICC/IF, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to residues in the N terminal of human NEDD8.

  • 阳性对照
    • Hela and K562 cell lysates and human lung carcinoma.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Alternative versions available:
    Anti-NEDD8 antibody (Alexa Fluor® 647) [Y297] (ab200849)

    Anti-NEDD8 antibody (Alexa Fluor® 488) [Y297] (ab201249)

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. Stable for 12 months at -20°C.
  • 存储溶液PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
  • Concentration information loading...
  • 纯度Protein A purified
  • 克隆单克隆
  • 克隆编号Y297
  • 同种型IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab81264 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 - 1/10000. Detects a band of approximately 9 kDa (predicted molecular weight: 9 kDa).
IHC-P 1/100 - 1/500.
ICC/IF 1/100 - 1/250.
IP 1/20 - 1/50.

靶标

  • 功能Ubiquitin-like protein which plays an important role in cell cycle control and embryogenesis. Covalent attachment to its substrates requires prior activation by the E1 complex UBE1C-APPBP1 and linkage to the E2 enzyme UBE2M. Attachment of NEDD8 to cullins activates their associated E3 ubiquitin ligase activity, and thus promotes polyubiquitination and proteasomal degradation of cyclins and other regulatory proteins.
  • 组织特异性Highly expressed in heart, skeletal muscle, spleen, thymus, prostate, testis, ovary, colon and leukocytes.
  • 序列相似性Belongs to the ubiquitin family.
  • 翻译后修饰Cleavage of precursor form by UCHL3 or SENP8 is necessary for function.
  • 细胞定位Nucleus. Mainly nuclear.
  • Information by UniProt
  • 数据库链接
  • 别名
    • FLJ43224 antibody
    • MGC104393 antibody
    • MGC125896 antibody
    • MGC125897 antibody
    • NED8 antibody
    • NEDD 8 antibody
    • NEDD-8 antibody
    • Nedd8 antibody
    • NEDD8_HUMAN antibody
    • Neddylin antibody
    • Neural precursor cell expressed developmentally down regulated 8 antibody
    • Neural precursor cell expressed developmentally down regulated gene 8 antibody
    • Neural precursor cell expressed developmentally down-regulated protein 8 antibody
    • Rub1 antibody
    • Ubiquitin like protein Nedd 8 antibody
    • Ubiquitin like protein Nedd8 antibody
    • Ubiquitin-like protein Nedd8 antibody
    see all

Anti-NEDD8 antibody [Y297] 图像

  • Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution (purified) + PC-12 whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 9 kDa
    Observed band size : 12,90 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

    The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.

  • Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution (purified) + K562 whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 9 kDa
    Observed band size : 12,90 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

    The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.

  • Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution (purified) + HEK293 cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size : 9 kDa
    Observed band size : 12,90 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

    The band at 12kDa is free NEDD8, and at 90kDa is Neddylated cullins.

  • ab81264 (purified) at 1/20 immunoprecipitating NEDD8 in 10 μg K562 cell lysate (Lanes 1 and 2, observed at 10 and 90 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
  • Immunofluorescence staining of HeLa cells with purified ab81264 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab81264 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab81264 at a working dilution of 1/400. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • All lanes : Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution (Unpurified)

    Lane 1 : C6 Cell Lysate
    Lane 2 : Raw264.7 Cell Lysate
    Lane 3 : PC-12 Cell Lysate
    Lane 4 : 3T3 Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size : 9 kDa
  • All lanes : Anti-NEDD8 antibody [Y297] (ab81264) at 1/500 dilution (Unpurified)

    Lane 1 : HeLa cell lysate
    Lane 2 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size : 9 kDa
    Observed band size : 9 kDa
    Other Bands: 90kDa Neddylated cullins
  • Western blot analysis of immunoprecipitation pellet from HeLa cell lysate using unpurified ab81264 at 1/500 dilution. Band at 90 kDa shows Neddylated cullins, 50 kDa the IgG fraaction and 9 kDa Free NEDD8.

  • Unpurified ab81264, at 1/100 dilution, staining NEDD8 in human lung carcinoma tissue by Immunohistochemistry using formalin-fixed, paraffin-embedded section.

Anti-NEDD8 antibody [Y297] (ab81264)参考文献

This product has been referenced in:
  • Lu R  et al. COPS5 amplification and overexpression confers tamoxifen-resistance in ERa-positive breast cancer by degradation of NCoR. Nat Commun 7:12044 (2016). Human . Read more (PubMed: 27375289) »
  • Godbersen JC  et al. The Nedd8-activating enzyme inhibitor MLN4924 thwarts microenvironment-driven NF-?B activation and induces apoptosis in chronic lymphocytic leukemia B cells. Clin Cancer Res 20:1576-89 (2014). Read more (PubMed: 24634471) »

See all 3 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Zebrafish Tissue lysate - whole (whole embryo)
Gel Running Conditions Non-reduced Denaturing (12%)
Loading amount 40 µg
Specification whole embryo
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Apr 06 2016

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