Skip to main content

Rabbit Recombinant Monoclonal NEDD8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Rat, Mouse samples. Cited in 42 publications.


Images

Western blot - Anti-NEDD8 antibody [Y297] (AB81264), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD8 antibody [Y297] (AB81264), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-NEDD8 antibody [Y297] (AB81264), expandable thumbnail
  • Immunoprecipitation - Anti-NEDD8 antibody [Y297] (AB81264), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-NEDD8 antibody [Y297] (AB81264), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 49% PBS, 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBICC/IFFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Expected
Expected
Rat
Expected
Tested
Expected
Expected
Expected

Tested
Tested

Species

Human

Dilution info

1/20 - 1/50

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Rat

Dilution info

1/1000 - 1/10000

Notes

-

Species

Mouse

Dilution info

1/1000 - 1/10000

Notes

-

Species

Human

Dilution info

1/1000 - 1/10000

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/300

Notes

-

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/500

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

2 products for Alternative Product

Target data

Function

Ubiquitin-like protein which plays an important role in cell cycle control and embryogenesis via its conjugation to a limited number of cellular proteins, such as cullins or p53/TP53 (PubMed:10318914, PubMed:10597293, PubMed:11953428, PubMed:14690597, PubMed:15242646, PubMed:9694792, PubMed:38605244). Attachment of NEDD8 to cullins is critical for the recruitment of E2 to the cullin-RING-based E3 ubiquitin-protein ligase complex, thus facilitating polyubiquitination and proteasomal degradation of cyclins and other regulatory proteins (PubMed:10318914, PubMed:10597293, PubMed:11953428, PubMed:20688984, PubMed:9694792, PubMed:38605244). Attachment of NEDD8 to p53/TP53 inhibits p53/TP53 transcriptional activity (PubMed:15242646). Covalent attachment to its substrates requires prior activation by the E1 complex UBE1C-APPBP1 and linkage to the E2 enzyme UBE2M (PubMed:14690597).

Alternative names

Recommended products

Rabbit Recombinant Monoclonal NEDD8 antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Rat, Mouse samples. Cited in 42 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

Y297

Purification technique

Affinity purification Protein A

Specificity

This antibody detects NEDD8 bound to cullins proteins (neddylated cullins) as well as free NEDD8.

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

Notes

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 monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

NEDD8 also known as neural precursor cell expressed developmentally downregulated protein 8 is a small ubiquitin-like protein with a molecular weight of approximately 9 kDa. It is expressed in various tissues prominently in the brain and muscles. NEDD8 conjugates to target proteins modifying their functions by altering their stability activity or location. It plays a role in the regulation of protein degradation.

Biological function summary

The NEDD8 protein is part of a modification system that activates cullin-RING ligase complexes. This activation promotes the ubiquitination and subsequent proteasomal degradation of targeted proteins. As a post-translational modification system similar to ubiquitin NEDD8 is important for maintaining protein homeostasis by ensuring timely degradation and turnover of cellular proteins. These processes impact cell cycle progression signal transduction and stress responses.

Pathways

NEDD8 is a central player in the ubiquitin-proteasome system an essential pathway for protein turnover. It is particularly important in the NEDDylation pathway where it regulates protein functions by transiently attaching to substrate proteins. Through these processes NEDD8 interacts closely with proteins like cullins and ubiquitin affecting cellular responses and maintaining cellular integrity under different conditions.

Associated diseases and disorders

NEDD8 influences several pathological states. Aberrant NEDD8 conjugation can lead to cancer by disrupting the controlled degradation of proteins involved in cell cycle regulation such as p27 and p21. Additionally improper NEDDylation has links to neurodegenerative diseases like Alzheimer's as it affects proteins related to synaptic function and neuronal survival. Understanding the role of NEDD8 in these diseases aids in the development of potential therapeutic strategies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

11 product images

  • Western blot - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Western blot - Anti-NEDD8 antibody [Y297] (ab81264)

    All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution

    Lane 1: C6 Cell Lysate at 20 µg

    Lane 2: Raw264.7 Cell Lysate at 20 µg

    Lane 3: PC-12 Cell Lysate at 20 µg

    Lane 4: 3T3 Cell Lysate at 20 µg

    Secondary

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

    Predicted band size: 9 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD8 antibody [Y297] (ab81264)

    Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab81264 at a working dilution of 1/400. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-NEDD8 antibody [Y297] (ab81264)

    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 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), used at a dilution of 1/1000. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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 (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at a dilution of 1/500. For negative control 2, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at a dilution of 1/400.

  • Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

    All lanes: Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)

    Predicted band size: 9 kDa

  • Flow Cytometry (Intracellular) - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-NEDD8 antibody [Y297] (ab81264)

    Intracellular Flow Cytometry analysis of A-673 (Human muscle Ewing's Sarcoma) cells labeling NEDD8 (red) with purified ab81264 at a 1/300 dilution (1ug/mL). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti rabbit IgG (Alexa Fluorr®488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (Black) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). Blue (unlabeled control) - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)

    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.

    All lanes: Immunoprecipitation - Anti-NEDD8 antibody [Y297] (ab81264)

    Predicted band size: 9 kDa

  • Western blot - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Western blot - Anti-NEDD8 antibody [Y297] (ab81264)

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

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

    All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution

    All lanes: PC-12 whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 9 kDa

    Observed band size: 12 kDa, 90 kDa

  • Western blot - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Western blot - Anti-NEDD8 antibody [Y297] (ab81264)

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

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

    All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution

    All lanes: K562 whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 9 kDa

    Observed band size: 12 kDa, 90 kDa

  • Western blot - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Western blot - Anti-NEDD8 antibody [Y297] (ab81264)

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

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

    All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/1000 dilution

    All lanes: HEK293 cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 9 kDa

    Observed band size: 12 kDa, 90 kDa

  • Western blot - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Western blot - Anti-NEDD8 antibody [Y297] (ab81264)

    Other Bands: 90kDa Neddylated cullins

    All lanes: Western blot - Anti-NEDD8 antibody [Y297] (ab81264) at 1/500 dilution

    Lane 1: HeLa cell lysate at 10 µg

    Lane 2: K562 cell lysate at 10 µg

    Secondary

    All lanes: HRP labelled goat anti-rabbit at 1/2000 dilution

    Predicted band size: 9 kDa

    Observed band size: 9 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD8 antibody [Y297] (ab81264), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NEDD8 antibody [Y297] (ab81264)

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

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com